Traditional in vitro strategies for sustainable production of bioactive compounds and manipulation of metabolomic profile in medicinal, aromatic and ornamental plants.
Precursor feeding, elicitation and culture medium parameters are traditional in vitro strategies to enhance bioactive compounds of medicinal, aromatic, and ornamental plants (MAOPs). Machine learning can help researchers find the best combination of these strategies to increase the secondary metabolites content of MAOPs. Many requirements for human life, from food, pharmaceuticals and cosmetics to clothes, fuel and building materials depend on plant-derived natural products. Essential oils, methanolic and ethanolic extracts of in vitro undifferentiated callus and organogenic cultures of medicinal, aromatic, and ornamental plants (MAOPs) contain bioactive compounds that have several applications for various industries, including food and pharmaceutical. In vitro culture systems provide opportunities to manipulate the metabolomic profile of MAOPs. Precursors feeding, elicitation and culture media optimization are the traditional strategies to enhance in vitro accumulation of favorable bioactive compounds. The stimulation of plant defense mechanisms through biotic and abiotic elicitors is a simple way to increase the production of secondary metabolites in different in vitro culture systems. Different elicitors have been applied to stimulate defense machinery and change the metabolomic profile of MAOPs in in vitro cultures. Plant growth regulators (PGRs), stress hormones, chitosan, microbial extracts and physical stresses are the most applied elicitors in this regard. Many other chemical tolerance-enhancer additives, such as melatonin and proline, have been applied along with stress response-inducing elicitors. The use of stress-inducing materials such as PEG and NaCl activates stress tolerance elicitors with the potential of increasing secondary metabolites content of MAOPs. The present study reviewed the state-of-the-art traditional in vitro strategies to manipulate bioactive compounds of MAOPs. The objective is to provide insights to researchers involved in in vitro production of plant-derived natural compounds. The present review provided a wide range of traditional strategies to increase the accumulation of valuable bioactive compounds of MAOPs in different in vitro systems. Traditional strategies are faster, simpler, and cost-effective than other biotechnology-based breeding methods such as genetic transformation, genome editing, metabolic pathways engineering, and synthetic biology. The integrate application of precursors and elicitors along with culture media optimization and the interpretation of their interactions through machine learning algorithms could provide an excellent opportunity for large-scale in vitro production of pharmaceutical bioactive compounds.


UV-Vis detection of hydrogen peroxide using horseradish peroxidase/copper phosphate hybrid nanoflowers.
A colorimetric platform based on enzyme-inorganic hybrid nanoflowers was used for ultraviolet-visible detection of hydrogen peroxide (H2O2). The hybrid nanoflowers were prepared via incubation at room temperature using horseradish peroxidase (HRP) and copper phosphate (Cu3(PO4)2) as the organic and inorganic components, respectively. HRP/Cu3(PO4)2 hybrid nanoflowers (HRP-HNFs) showed linear H2O2 detection range from 100 nM to 100 μM, moreover, HRP-HNFs exhibited good selectivity for H2O2 detection due to the specificity of HRP. In addition, HRP-HNFs have good reusability, excellent color stability and long-term storage stability, which indicate HRP-HNFs are promising for catalysis and biomedical applications.


Pain, motor and gait assessment of murine osteoarthritis in a cruciate ligament transection model.
The major complaint of Osteoarthritis (OA) patients is pain. However, due to the nature of clinical studies and the limitation of animal studies, few studies have linked function impairment and behavioral changes in OA animal models to cartilage loss and histopathology. Our objective was to study surrogate markers of functional impairment in relation to cartilage loss and pathological changes in a post-traumatic mouse model of OA. We performed a battery of functional analyses in a mouse model of OA generated by cruciate ligament transection (CLT). The changes in functional analyses were linked to histological changes graded by OARSI standards, histological grading of synovitis, and volumetric changes of the articular cartilage and osteophytes quantified by phase contrast micro-computed tomography (μCT). OA generated by CLT led to decreased time on rotarod, delayed response on hotplate analysis, and altered gait starting from 4 weeks after surgery. Activity in open field analysis did not change at 4, 8, or 12 weeks after CLT. The magnitude of behavioral changes was directly correlated with higher OARSI histological scores of OA, synovitis in the knee joints, cartilage volume loss, and osteophyte formation. Our findings link functional analyses to histological grading, synovitis, comprehensive three-dimensional assessment of cartilage volume and osteophyte formation. This serves as a reference for a mouse model in predicting outcomes of OA treatment.


A developmental study of timing behavior in 4 1/2- and 7-year-old children.
The present study investigated effects of age and instructions on temporal regulations of behavior in children. In the first experiment 4 1/2-year-old and 7-year-old subjects were trained with a DRL (differential reinforcement of low rates) 5-s and a DRL 10-s schedule. Results demonstrate that age and timing performance are related. Seven-year-olds are more efficient than the 4 1/2-year-olds. A striking decline in the 4 1/2-year-old children's capacity to space responses was observed in the DRL 10-s schedule as compared to the DRL 5-s schedule. Analysis of individual performances suggests that the evolution of DRL performance between 4 and 7 years of age depends not only on the development of the capacity to delay responding but also on the acquisition of the ability to represent the reinforcement contingencies, that is, the temporal parameters of the task to oneself. In order to test this hypothesis a second experiment was conducted where instructions to wait between operant responses were given to a group of 4 1/2-year-old subjects at the beginning of a DRL 5-s and a DRL 10-s schedule. The results show that these instructions enhance DRL performance. By directing the 4 1/2-year-old subjects' attention to the temporal requirements of the task, instructions led to efficient performance and accurate timing of responses to the DRL schedule.


Stability of synaptic density and spine volume in dentate gyrus of aged rats.
The number of synapses per unit volume and per granule cell and the size of dendritic spines were studied in the dentate gyrus of Sprague-Dawley rats 6, 24, and 30 months of age. Neither synaptic density nor mean spine volume showed any age-related trends. An increase in granule cell packing density at 24 months and concomitant stability of the height of the granule cell layer is consistent with the idea that postnatal generation of granule cells may continue late into life. Possible explanations for the discrepancies in the literature regarding synaptic loss in this area include differences in morphometric techniques, age of animals used, regional differences within dentate gyrus, and sampling variability. Generalized synapse loss in the senescent rodent brain remains to be established.


Biochemical characteristics of a textile dye degrading extracellular laccase from a Bacillus sp. ADR.
Bacillus sp. ADR secretes an extracellular laccase in nutrient broth, and this enzyme was purified up to 56-fold using acetone precipitation and DEAE-cellulose anion exchange chromatography. The molecular weight of purified laccase was estimated to be 66 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified laccase oxidized 2,6-dimethoxy phenol, o-tolidine, hydroquinone, L-DOPA and guaiacol. The optimum pH for oxidation of o-tolidine, 2,6-dimethoxy phenol and guaiacol were 3.0, 4.0 and 5.0, respectively. The purified laccase contained 2.7 mol/mol of copper. The laccase was stable up to 40 °C and within the pH range of 7.0-9.0. Well-known inhibitors of multicopper oxidases such as, sodium azide, L-cysteine and dithiothreitol showed significant inhibition of laccase activity. The purified enzyme decolorized structurally different azo dyes with variable decolorization rates and efficiencies of 68-90%. This study is useful for understanding the precise use of Bacillus sp. ADR in the decolorization of textile dyes containing industrial wastewater.


Interrelationships among trace metals and metallothionein in digestive glands and gills for field samples of Merceneria merceneria.
More widespread use of metallothionein (MT) as a biomarker for trace metal pollution continues to be partly dependent on obtaining reliable baseline concentrations and identifying increased induction of the enzyme with only modest increases in metal concentrations. In this study, new data on metals and MT levels in whole clams tissue, gills, and digestive glands from field samples and in sediments are presented. Concentrations of Cd, Cu, Fe, and Zn in depurated (24 h) clam samples of digestive glands, gills, and the whole clam Merceneria merceneria from the Indian River Lagoon, Florida, varied with location and showed moderate to strong correlations among Zn, Cu, and Fe. Concentrations of metallothionein (dry wt.) ranged from 34─270 μg/g in gills and 150-440 μg/g in digestive glands and showed moderate to strong correlations between organs and with metal concentrations in those organs. Observed trends support increased synthesis of metallothionein with only moderate increases in metal values and in response to statistically higher sediment metal concentrations.


Successful treatment of parathyroid cyst using ethanol sclerotherapy.
Simple parathyroid cysts are rare; only about 200 cases have been reported in the literature. The diagnosis was facilitated recently by parathyroid hormone assay of the cyst fluid. Standard treatment consists of percutaneous aspiration of the cyst and surgical excision for recurrent lesions. Sclerotherapy of recurrent lesions was described in two cases and results were promising. The authors report a new case of recurrent nonfunctioning parathyroid cyst in a 34-year-old female, with complete resolution after ethanol sclerosis and no recurrence for 15 months of follow-up. Ethanol sclerotherapy is recommended as a simple and safe treatment for recurrent parathyroid cysts.


Strains of Streptococcus pyogenes from severe invasive infections bind HEp2 and HaCaT cells more avidly than strains from uncomplicated infections.
Epidemiologically unrelated Streptococcus pyogenes strains isolated from blood, throat, and skin were assayed for adherence to HEp2 and HaCaT cells. Invasive isolates showed significantly higher avidity for these cell lines than isolates from skin and throat. In general, S. pyogenes showed greater binding to HaCaT cells than to HEp2 cells.


[Effect of petroleum on the function and morphological picture of the liver].
The male rats of Wistar strain were exposed to the vapours of arabic petroleum Abu-Dhaki Zakun at concentration of 1500 mg/m3 during 12 weeks, 6 times weekly per 90 minutes daily. The biochemical examinations of blood serum (aminotransferase, alkaline phosphatase total protein level and its fractions, bilirubin level) and histopathological and histochemical examinations of the liver have revealed no deviations from the normal state.


Nutritional screening--evaluation and implementation of a simple Nutrition Risk Score.
Development of a simple, validated Nutrition Risk Score for identification of patients at risk of undernutrition is described. The score is easy to use, applicable to all patient categories and ages, and correlated well with a validated Nutrition Risk Index (r = 0.68, p < 0.001) and clinical impression (r = 0.83, p < 0.001). Reproducible scores were obtained between dietitians (r = 0.91, p < 0.001) and between dietitians and nursing staff (r = 0.80, p < 0.001). The score was used to assess risk of undernutrition in 153 consecutive admissions to medical and surgical specialties. Patients were categorised as low (50%), moderate (24%) or high risk (26%). Evaluation of measures to prevent nutritional depletion revealed that no action was taken in 64% (23/36) of moderate risk and 30% (12/40) of high risk patients. Implementation of routine, hospital-wide nutritional screening, to identify patients in need of nutritional support, has been achieved by inclusion of the score in the standard nursing assessment process.


Useful application of negative suction drainage on the umbilicus after transaxillary breast augmentation.
Simple application of negative suction drains through the umbilicus during transaxillary breast augmentation obtained a more natural drainage and an easier positioning of the drain at the dependant portions than placement of the drains at the transaxillary incision site. Moreover, the patient was more satisfied due to increased comfort while wearing clothes, which resulted in a quicker recovery to everyday activities. In addition, the scar of the drain site was hidden on the inner side of the umbilicus. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors at www.springer.com/00266.


Low vitamin D levels are associated with increased rejection and infections after lung transplantation.
The prevalence of vitamin D deficiency in lung disease is greater than in the general population. Vitamin D deficiency may negatively affect immune and lung function. Accordingly, we hypothesized that lung transplant recipients with vitamin D deficiency are more susceptible to rejection and infections after transplantation. Transplant outcomes were reviewed in a retrospective cohort of 102 lung transplant recipients who had 25-hydroxyvitamin D [25(OH)D] levels drawn during the near-transplant period (100 days pre- or post-transplant). In the near-transplant period, 80% of recipients were 25(OH)D-deficient and 20% were not 25(OH)D-deficient. Episodes of acute cellular rejection in the deficient group were more frequent than in the non-deficient group [mean 1.27 (0.99 to 1.55) vs 0.52 (0.12 to 0.93), p = 0.006]. The rejection rate in the deficient group was more than double that of the the non-deficient group [IRR 2.43 (1.30 to 4.52), p = 0.005]. Infectious episodes were also more frequent in the deficient group than in the non-deficient group [mean 4.01 (3.24 to 4.79) vs 2.71 (1.47 to 3.96), p = 0.04]. The mortality rate of recipients who remained 25(OH)D-deficient 1 year after transplant was almost 5-fold higher than in recipients who were not 25(OH)D-deficient [IRR 4.79 (1.06 to 21.63), p = 0.04]. Low serum 25(OH)D levels in lung transplant recipients were associated with increased incidence of acute rejection and infection. The mortality of recipients who remained deficient 1 year post-transplant was higher than that of recipients who maintained normal vitamin D levels at 1 year post-transplant.


miR-223-3p targets FBXW7 to promote epithelial-mesenchymal transition and metastasis in breast cancer.
Breast cancer is the most common malignant tumor diagnosed in women. It is the second leading cause of cancer-related death among women in the world. Aberrant expression of microRNAs (miRNAs) have been identified to be involved in the development and progression of breast cancer. The aim of this study was to investigate the function of miR-223-3p in breast cancer progression and metastasis. qRT-PCR was used to analyze the expression levels of miR-223-3p in breast cancer tissues and cell lines. Wound healing and Matrigel assays were used to examine cell motility and invasiveness. FBXW7 3'-UTR construct and luciferase reporter assays were performed for the target gene. miR-223-3p was overexpressed in breast cancer tissue and cell lines. A high level of miR-223-3p was associated with poor prognosis in breast cancer patients. In addition, overexpressed miR-223-3p promoted the migration and invasion of breast cancer cells in vitro and in vivo. Mechanistically, we found that tumor suppressor gene FBXW7 is a target of miR-223-3p. Luciferase activity reporter assay indicated miR-223-3p could directly bind with the 3'-UTR of FBXW7. miR-223-3p exhibited its oncogenic role partly by decreasing FBXW7 expression, and consequently promoted the invasion and metastasis of breast cancer cells. Our study revealed a physical and functional relationship among miR-223-3p and FBXW7. By negatively regulating FBXW7 expression, miR-223-3p exerts a tumor promotion role promoting cell invasion and metastasis in breast cancer.


Fetal anomaly diagnosis and termination of pregnancy in Ireland: a service evaluation following implementation of abortion services in 2019.
Approximately 3% of pregnancies are diagnosed with a fetal anomaly, of which a proportion is fatal or life-limiting. The introduction of legislation for termination of pregnancy in Ireland in 2019 for conditions "likely to lead to the death of the fetus" made termination of pregnancy for "fatal fetal anomaly" an option for pregnant women in Ireland. This study examined all cases of termination of pregnancy performed for major fetal anomaly over the first 3 years of service implementation, including cases that did not meet the legal criteria, resulting in women traveling outside Ireland for abortion care. A retrospective service evaluation of tertiary fetal medicine clinics in 2 tertiary maternity hospitals between 2019 and 2021 was undertaken. We compared pregnancies of patients diagnosed with fatal fetal anomaly who underwent termination of pregnancy in Ireland with those of patients who did not meet the legal criteria and hence traveled outside Ireland for termination of pregnancy. Overall, 139 pregnancies met the inclusion criteria; 83 (59.7%) patients had termination of pregnancy in the tertiary maternity hospital (local), and 56 (40.3%) traveled abroad, mainly to the United Kingdom. Demographic characteristics were similar between the 2 groups, as was gestation at diagnosis and delivery. All cases where termination of pregnancy was local were discussed at fetal medicine multidisciplinary meetings, as opposed to 34% of cases of patients who ultimately traveled outside Ireland for termination of pregnancy. The most common indication (25/83; 30.1%) for local termination of pregnancy was trisomy 18, followed by anencephaly. Traveling to obtain abortion care was mainly due to diagnosis of trisomy 21 (30/56; 53.6%), followed by other multiple structural anomalies/syndromes deemed locally as not meeting the legal criteria. Legislation for termination of pregnancy for fetal anomaly, restricted to fatal diagnoses, is difficult to implement, requires significant multidisciplinary input, and can lead to limited services for pregnancies diagnosed with major fetal anomalies. Our findings emphasize the impact of legislative barriers to abortion care for fetal anomaly and the need for policies and services that support women's access to termination of pregnancy for fetal anomaly.


Lack of interchangeability between visual analogue and verbal rating pain scales: a cross sectional description of pain etiology groups.
Rating scales like the visual analogue scale, VAS, and the verbal rating scale, VRS, are often used for pain assessments both in clinical work and in research, despite the lack of a gold standard. Interchangeability of recorded pain intensity captured in the two scales has been discussed earlier, but not in conjunction with taking the influence of pain etiology into consideration. In this cross-sectional study, patients with their pain classified according to its etiology (chronic/idiopathic, nociceptive and neuropathic pain) were consecutively recruited for self-assessment of their actual pain intensity using a continuous VAS, 0-100, and a discrete five-category VRS. The data were analyzed with a non-parametric statistical method, suitable for comparison of scales with different numbers of response alternatives. An overlapping of the VAS records relative the VRS categories was seen in all pain groups. Cut-off positions for the VAS records related to the VRS categories were found lower in patients with nociceptive pain relative patients suffering from chronic/idiopathic and neuropathic pain. When comparing the VAS records transformed into an equidistant five-category scale with the VRS records, systematic disagreements between the scales was shown in all groups. Furthermore, in the test-retest a low percentage of the patients agreed to the same pain level on the VAS while the opposite hold for the VRS. The pain intensity assessments on VAS and VRS are in this study, not interchangeable due to overlap of pain records between the two scales, systematic disagreements when comparing the two scales and a low percentage intra-scale agreement. Furthermore, the lower VAS cut-off positions relative the VRS labels indicate different meaning of the rated pain intensity depending on pain etiology. It is also indicated that the scales have non-linear properties and that the two scales probably have different interpretation. Our findings are in favor of using the VRS in pain intensity assessments but if still the VAS is preferred, the VAS data should be analyzed as continuous using statistical methods suitable for ordinal data. Furthermore, our findings indicate a risk to over or under estimate the patient's perceived pain when interpreting condensed VAS data.


Advances in Human Stem Cells and Genome Editing to Understand and Develop Treatment for Fragile X Syndrome.
Fragile X syndrome (FXS), the most common genetic form of autism spectrum disorder, is caused by deficiency of the fragile X mental retardation protein (FMRP). Despite extensive research using animal models, understanding how FMRP regulates human brain development and function remains a major challenge. Human pluripotent stem cells (hPSCs) offer powerful platforms for studying mechanisms of human diseases and for evaluating potential treatments. Genome editing, particularly the CRISPR/Cas9-based method, is highly effective for generating models to study genetic human diseases. Here we summarize how hPSCs and genome editing provide much-needed models for studying the genetic underpinnings, cellular mechanisms, and neuropathology that are unique to human FXS. The use of hPSCs and genome editing also provides an essential platform for therapeutic development in FXS.


Facebook as a Medium for the Support and Enhancement of Ambulatory Care for People With Diabetes: Qualitative Realist Evaluation of a Real-World Trial.
There is a growing focus on the potential uses, benefits, and limitations of social media in the context of health care communication. In this study, we have sought to evaluate an initiative pioneered at a hospital in Denmark that uses Facebook to support and enhance patient-provider communication about diabetes. This paper aims to evaluate the success of the trial according to its initial objectives and to assess its potential scalability. The study was undertaken in a clinic for diabetes and hormonal diseases at a large regional hospital in Denmark. Using a realist evaluation approach, we identified 4 key components in the program theory of the initiative, which we formulated as context-mechanism-outcome configurations (eg, complex and iterative chains of causality). These configurations informed data gathering and analysis. Primary data sources were the activity and content in the Facebook group, in the form of posts, likes, and comments, and interviews with patients (n=26) and staff (n=6) at the clinic. New developments in diabetes technology were the most popular posts in the forum, judged by number of likes and comments. Otherwise, information specific to the clinic received the most attention. All 4 components of the program theory were compromised to varying degrees, either as a result of failings in the anticipated mechanisms of change or contextual factors derived from the mode of implementation. Social media serves well as a conduit for imagining positive change, but this can be a strength and weakness when attempting to enact change via concrete interventions, where stakeholder expectations may be unreasonably high or incompatible. Nonetheless, such initiatives may possess intangible benefits difficult to measure in terms of cost-effectiveness.


Cassava postharvest physiological deterioration: a complex phenomenon involving calcium signaling, reactive oxygen species and programmed cell death.
Postharvest physiological deterioration (PPD) of cassava (Manihot esculenta) storage roots is a complex physiological and biochemical process which involve many regulatory networks linked with specific proteins modulation and signaling transduction pathways. However, it is poorly understood regarding biological regulation, and the interactions among protein groups and signals to determine PPD syndrome in cassava storage roots. This review sheds some light on the possible molecular mechanisms involved in reactive oxygen species (ROS), calcium signaling transduction, and programmed cell death (PCD) in cassava PPD syndrome. A model for predicting crosstalk among calcium signaling, ROS and PCD is suggested to fine-tune PPD syndrome. This would clues to cassava molecular breeding to alleviate the PPD effects on the shelf-life.


Tacrine slows the rate of ageing of sarin-inhibited acetylcholinesterase.
Bovine erythrocyte acetylcholinesterase was inhibited by the organophosphate sarin, and the rate of ageing (the time-dependent decrease in the ability of an oxime to reactivate the enzyme) was studied. At pH 7.0 and 37 degrees C, 10(-5) M or 10(-6) M tacrine (tetrahydroaminoacridine) decreased the rate of ageing in low ionic strength buffer. Tacrine at 10(-5) M also significantly decreased the rate of ageing in 150 mM NaCl. The results indirectly demonstrated that the inhibition of substrate hydrolysis by tacrine is reversible, and that tacrine does not prevent reactivation of sarin-inhibited acetylcholinesterase. Both these observations, which were also made for rat brain acetylcholinesterase, are in contrast with reports in the literature.


Detailed and abstract phase-locked attractor network models of early olfactory systems.
Across species, primary olfactory centers show similarities both in their cellular organization and their types of olfactory information coding. In this article, we consider an excitatory-inhibitory spiking neural network as a model of early olfactory systems (antennal lobe for insects, olfactory bulb for vertebrates). In line with experimental results, we show that, in our network, odor-like stimuli evoke synchronization of excitatory cells, phase-locked to the oscillations of the local field potential. As revealed by a mathematical analysis, the phase-locking probability of excitatory cells is given by an inverted-U function and the firing probability of inhibitory cells is well described by a sigmoid function. These neural response functions are used to reduce the spiking model to a more abstract model with discrete-time dynamics (oscillatory cycles) and binary-state neurons (phase-locked or not). An iterative map, built for explaining the dynamics of the binary model, reveals that it converges to fixed point attractors similar to those obtained with the spiking model. This result is consistent with odor-specific attractors found in recent experimental studies. It also provides insights for designing bio-inspired olfactory associative memories applicable for data analysis in electronic noses.


Asymptomatic latent syphilis in geriatric patients in association with stoke: Analysis using routine syphilis tests.
We investigated the association between stroke and asymptomatic latent syphilis (ALS) in geriatric patients. This retrospective observational study included patients aged >65 years who underwent routine qualitative rapid plasma reagin (RPR) and Treponema pallidum Latex Agglutination (TPLA) tests at a 161-bed community acute care hospital with long-term care facilities in Kamakura, Japan, from August 2014 to February 2019. Asymptomatic patients with a positive TPLA test result were diagnosed with ALS. Ninety-six patients with ALS were included in the study. Fifty-one patients (53.1%) had a positive RPR test. Comorbidities included hypertension (n = 44; 45.8%), chronic kidney disease (n = 44; 45.8%), and fracture (n = 29; 30.2%). No significant differences were found in sex, age, or comorbidities in univariate analyses. Multivariate analysis of the TPLA-positive geriatric patients revealed that a positive RPR test (odds ratio = 9.06; 95% confidence interval = 1.69-48.5; p = 0.01) was associated with a history of stroke. For geriatric patients with ALS, a positive qualitative RPR test was associated with a history of stroke. Medical evaluation and management of the risk factors for stroke are more necessary for ALS with a positive RPR qualitative test.


Stenosis after stapler anastomosis.
Two cases of anastomotic stenosis after the use of the GIA Auto Suture Stapler are presented as examples of the potential problem that does exist in using this instrument. Possible causes and a suggestion for eliminating this complication have been outlined.


Effects of vitreomacular adhesion on anti-vascular endothelial growth factor treatment for polypoidal choroidal vasculopathy.
To evaluate the effect of posterior vitreomacular adhesion (VMA), documented by optical coherence tomography, on the outcome of anti-vascular endothelial growth factor treatment of polypoidal choroidal vasculopathy. Medical records of 102 patients (104 eyes) with polypoidal choroidal vasculopathy were retrospectively reviewed and categorized according to the presence of posterior VMA into 2 subgroups: VMA positive (+) group (23 eyes) and VMA negative (-) group (81 eyes). Best-corrected visual acuity and central macular thickness after antivascular endothelial growth factor treatment were compared between the 2 groups at baseline and at 1 month, 3 months, 6 months, and 12 months. At the last follow-up, average number of injections was 4.82 ± 1.27 in the VMA (+) group and 4.92 ± 1.45 in the VMA (-) group. After injection, the mean logarithm of the minimum angle of resolution of best-corrected visual acuity improved from 0.81 ± 0.53 (Snellen equivalent, 20/129) to 0.67 ± 0.52 (Snellen equivalent, 20/93) in the VMA (+) group (P = 0.01) and from 0.79 ± 0.50 (Snellen equivalent, 20/123) to 0.64 ± 0.58 (Snellen equivalent, 20/91) in the VMA (-) group (P = 0.02). Average central macular thickness decreased from 354.4 ± 124.5 μm to 249.6 ± 112.5 μm in the VMA (+) group (P = 0.01) and from 361.2 ± 140.2 μm to 267.3 ± 103.5 μm in the VMA (-) group (P = 0.01). Polyp regression rate was 21.7% (5 eyes of 23 eyes) in the VMA (+) group and 22.2% (18 eyes of 81 eyes) in the VMA (-) group. There was no statistically significant difference in the best-corrected visual acuity improvement, central macular thickness improvement, and polyp regression rate between the groups. Unlike typical age-related macular degeneration, posterior VMA was not associated with a visual outcome after intravitreal antivascular endothelial growth factor for polypoidal choroidal vasculopathy.


Dose-rate effects of ethyl methanesulfonate on survival and mutation induction in cultured Chinese hamster cells.
The dose-rate effects of ethyl methanesulfonate (EMS) on the survival and induction of mutations in Chinese hamster Don cells were investigated. The most effective time of exposure to EMS for reducing the surviving fraction of cells was 4 h, shorter and longer exposure times being less effective. The threshold or minimal concentration of EMS giving a surviving fraction of 0.5 was 0.05 mg/ml. The minimal effective time of exposure to EMS for cell death was 1 h. Corrected survival curves showed that longer exposure times at lower dose rates of EMS had less cytotoxic effect than shorter exposure times at higher dose rates. After exposure of Don cells to various doses of EMS for various times, the frequencies of mutations resistant to 6--thioguanine (6TG) were measured. An exposure time of 4 h produced a lower mutation frequency than shorter or longer exposure times that resulted in the same surviving fraction of cells. An exposure time of 20 h produced the highest induced mutation frequency. This system using cultured Chinese hamster cells should be useful as a sensitive procedure for detecting the mutagenic actions of chemicals.


Lymphocyte markers in the bovine foetus.
The ontogeny of lymphocyte surface markers was studied in thymus, retropharyngeal lymph nodes, spleen, bone marrow, and liver of 48 bovine foetuses, 80-280 days of gestation. The markers examined were the spontaneous (E) rosette against sheep erythrocytes (SRBCs) in foetal calf serum (E/FCS), in medium supplemented with gelatin (E/CFG), and in medium supplemented with dextran (E/dextran), the erythrocyte-antibody (EA) rosette for immunoglobulin (Fc) receptors, the erythrocyte-antibody-complement (EAC) rosette for complement receptors, and the immunofluorescence test (IFT) for surface-membrane immunoglobulins (SmIg). E-rosette-forming cells (RFCs) occurred in the thymus in substantial numbers throughout the period studied, in spleen and lymph nodes only after 160 days, and in bone marrow and liver rarely. The dynamics of E rosettes formed in the three supplements suggested that each supplement might be supporting rosette formation by different populations of cells, albeit considerable overlap might be expected. Organ distribution and development of EA and EAC RFCs did not parallel the expected ontogeny of any class of lymphocytes. Cells carrying SmIg developed mainly in spleen and lymph nodes. Towards the end of gestation some cells in liver also had SmIg.


Thoracic outlet syndrome.
Eleven patients with vascular sequelae of thoracic outlet syndrome were operated on at the University of California, SanFrancisco, during the past 17 years. Five patients presented with episodes of ischemia of the arm and hand secondary to microemboli released from subclavian arterial lesions produced by chronic compression at the thoracic outlet. Treatment consisted of arterial reconstruction, removal of the compressive structure, and cervical sympathectomy to relieve or lessen distal ischemia. Four of the five patients had good or excellent results; one patient required amputation of the forearm. The results were inversely proportional to the extent of distal arterial embolic occlusions present at the time of surgical treatment. Six patients presented with symptoms of chronic venous hypertension. Four of the six had subclavian venous thrombosis and were treated by transaxillary resection of the first rib to decompress the collateral veins within the costoclavicular space. All four were symptomatically improved. Two patients had venous hypertension due to extrinsic compression of the subclavian vein. One patient became asymptomatic and the other was markedly improved after resection for external compression. In this small series transaxillary resection of the first rib has resulted in symptomatic improvement in chronic venous hypertension of the arm.


Sigmoid volvulus in adolescence.
Sigmoid volvulus in young adults is rare. It usually presents as chronic recurrent attacks of abdominal pain. The diagnosis is usually delayed because of a low level of suspicion. We present the case of a 17-year-old girl with recurrent episodes of sigmoid volvulus, to illustrate the problem.


Sleep-wake behavior and responses to sleep deprivation of mice lacking both interleukin-1 beta receptor 1 and tumor necrosis factor-alpha receptor 1.
Data indicate that interleukin (IL)-1 beta and tumor necrosis factor-alpha (TNFalpha) are involved in the regulation of non-rapid eye movement sleep (NREMS). Previous studies demonstrate that mice lacking the IL-1 beta type 1 receptor spend less time in NREMS during the light period, whereas mice lacking the p55 (type 1) receptor for TNFalpha spend less time in NREMS during the dark period. To further investigate roles for IL-1 beta and TNFalpha in sleep regulation we phenotyped sleep and responses to sleep deprivation of mice lacking both the IL-1 beta receptor 1 and TNFalpha receptor 1 (IL-1R1/TNFR1 KO). Male adult mice (IL-1R1/TNFR1 KO, n=14; B6129SF2/J, n=14) were surgically instrumented with EEG electrodes and with a thermistor to measure brain temperature. After recovery and adaptation to the recording apparatus, 48 h of undisturbed baseline recordings were obtained. Mice were then subjected to 6h sleep deprivation at light onset by gentle handling. IL-1R1/TNFR1 KO mice spent less time in NREMS during the last 6h of the dark period and less time in rapid eye movement sleep (REMS) during the light period. There were no differences between strains in the diurnal timing of delta power during NREMS. However, there were strain differences in the relative power spectra of the NREMS EEG during both the light period and the dark period. In addition, during the light period relative power in the theta frequency band of the REMS EEG differed between strains. After sleep deprivation, control mice exhibited prolonged increases in NREMS and REMS, whereas the duration of the NREMS increase was shorter and there was no increase in REMS of IL-1R1/TNFR1 KO mice. Delta power during NREMS increased in both strains after sleep deprivation, but the increase in delta power during NREMS of IL-1R1/TNFR1 KO mice was of greater magnitude and of longer duration than that observed in control mice. These results provide additional evidence that the IL-1 beta and TNFalpha cytokine systems play a role in sleep regulation and in the alterations in sleep that follow prolonged wakefulness.


Simulations on the influence of myelin water in diffusion-weighted imaging.
While myelinated axons present an important barrier to water diffusion, many models used to interpret DWI signal neglect other potential influences of myelin. In this work, Monte Carlo simulations were used to test the sensitivity of DWI results to the diffusive properties of water within myelin. Within these simulations, the apparent diffusion coefficient (D app) varied slowly over several orders of magnitude of the coefficient of myelin water diffusion (D m), but exhibited important differences compared to D app values simulated that neglect D m (=0). Compared to D app, the apparent diffusion kurtosis (K app) was generally more sensitive to D m. Simulations also tested the sensitivity of D app and K app to the amount of myelin present. Unique variations in D app and K app caused by differences in the myelin volume fraction were diminished when myelin water diffusion was included. Also, expected trends in D app and K app with experimental echo time were reduced or inverted when accounting for myelin water diffusion, and these reduced/inverted trends were seen experimentally in ex vivo rat brain DWI experiments. In general, myelin water has the potential to subtly influence DWI results and bias models of DWI that neglect these components of white matter.


Coarctation of aorta combined with multiple aneurysms.
We reported a case of a 53-year-old patient with coarctation of the aorta and multiple aneurysmatic changes on the aortic arch. Enhanced computed tomography and reconstruction revealed significant coarctation and multiple aneurysmatic dilatations. The patient underwent stent implantation and was discharged with symptoms relieved. Follow-up examination progression of aneurysms, however, without symptoms.


A molecular pathway determining left-right asymmetry in chick embryogenesis.
While significant progress has been made in understanding the molecular events underlying the early specification of the antero-posterior and dorso-ventral axes, little information is available regarding the cellular or molecular basis for left-right (LR) differences in animal morphogenesis. We describe the expression patterns of three genes involved in LR determination in chick embryos: activin receptor IIa, Sonic hedgehog (Shh), and cNR-1 (related to the mouse gene nodal). These genes are expressed asymmetrically during and after gastrulation and regulate the expression of one another in a sequential pathway. Moreover, manipulation of the sidedness of either activin protein or Shh expression alters heart situs. Together, these observations identify a cascade of molecular asymmetry in that determines morphological LR asymmetry in the chick embryo.


Osteocytes directly regulate osteolysis via MYD88 signaling in bacterial bone infection.
The impact of bone cell activation on bacterially-induced osteolysis remains elusive. Here, we show that matrix-embedded osteocytes stimulated with bacterial pathogen-associated molecular patterns (PAMPs) directly drive bone resorption through an MYD88-regulated signaling pathway. Mice lacking MYD88, primarily in osteocytes, protect against osteolysis caused by calvarial injections of bacterial PAMPs and resist alveolar bone resorption induced by oral Porphyromonas gingivalis (Pg) infection. In contrast, mice with targeted MYD88 restoration in osteocytes exhibit osteolysis with inflammatory cell infiltration. In vitro, bacterial PAMPs induce significantly higher expression of the cytokine RANKL in osteocytes than osteoblasts. Mechanistically, activation of the osteocyte MYD88 pathway up-regulates RANKL by increasing binding of the transcription factors CREB and STAT3 to Rankl enhancers and by suppressing K48-ubiquitination of CREB/CREB binding protein and STAT3. Systemic administration of an MYD88 inhibitor prevents jawbone loss in Pg-driven periodontitis. These findings reveal that osteocytes directly regulate inflammatory osteolysis in bone infection, suggesting that MYD88 and downstream RANKL regulators in osteocytes are therapeutic targets for osteolysis in periodontitis and osteomyelitis.


Mutagenesis of surfactant protein D informed by evolution and x-ray crystallography enhances defenses against influenza A virus in vivo.
The recognition of influenza A virus (IAV) by surfactant protein D (SP-D) is mediated by interactions between the SP-D carbohydrate recognition domains (CRD) and glycans displayed on envelope glycoproteins. Although native human SP-D shows potent antiviral and aggregating activity, trimeric recombinant neck+CRDs (NCRDs) show little or no capacity to influence IAV infection. A mutant trimeric NCRD, D325A/R343V, showed marked hemagglutination inhibition and viral neutralization, with viral aggregation and aggregation-dependent viral uptake by neutrophils. D325A/R343V exhibited glucose-sensitive binding to Phil82 hemagglutinin trimer (HA) by surface plasmon resonance. By contrast, there was very low binding to the HA trimer from another virus (PR8) that lacks glycans on the HA head. Mass spectrometry demonstrated the presence of high mannose glycans on the Phil82 HA at positions known to contribute to IAV binding. Molecular modeling predicted an enhanced capacity for bridging interactions between HA glycans and D325A/R343V. Finally, the trimeric D325A/R343V NCRD decreased morbidity and increased viral clearance in a murine model of IAV infection using a reassortant A/WSN/33 virus with a more heavily glycosylated HA. The combined data support a model in which altered binding by a truncated mutant SP-D to IAV HA glycans facilitates viral aggregation, leading to significant viral neutralization in vitro and in vivo. These studies demonstrate the potential utility of homology modeling and protein structure analysis for engineering effective collectin antivirals as in vivo therapeutics.


Conformational characteristics of the dinucleoside triphosphate pCpGp from energy-minimization studies.
The influence of the 3'- and 5'- terminal phosphates on the conformational characteristics of the dinucleoside monophosphate CpG is described in this paper. The computed potential energy of the system is minimized with respect to the relevant 10 dihedral angles permitting the two sugar rings to adopt the alternative puckering states, 2E and 3E. Of the 84 conformations considered, 22 become energetically accessible. The familiar A-, B-, Z- and Watson-Crick-type backbone states of DNA subunits become low-energy forms for this RNA unit pCpGp also. The Watson-Crick-type backbone is invariably preferred in all the four sugar pucker sequences, indicating its importance in the dynamics of sugar pucker fluctuations and in the DNA-RNA association. The interphosphate geometries and the possible hydrogen-bonded states are discussed in relation to the varied folded/extended polynucleotide structures.


Is the cytoskeleton-plasma membrane complex involved in lens protein biosynthesis?
Calf lens fiber cells contain a population of polyribosomes that direct, at least in vitro, the synthesis of a specific plasma membrane protein MP26. This protein may serve as a marker in terminal differentiation, since it is absent in the lens epithelium but appears in lens fiber plasma membranes. The MP26 manufacturing polyribosomes are found to be associated with a structural complex in which also the cytoskeleton and plasma membranes participate. They can be released from the complex by treatment with DNAse I. This result presumably reflects the involvement of actin in the linkage of the MP26 synthesizing polyribosomes to the cytoskeleton-membrane complex.


Mechanisms of endothelin-1-induced pulmonary vasodilatation in neonatal pigs.
1. We determined the contributions of three independent vasodilator mechanisms (cyclo-oxygenase metabolites, nitric oxide and ATP-sensitive potassium channels) in the mediation of pulmonary vasomotor effects of endothelin-1 (ET-1) in neonatal pigs. 2. Lungs of piglets (2.7 +/- 0.3 days old) were perfused at constant flow (60 ml min-1) with recirculating Ringer-albumin solution. We measured pulmonary artery pressure (Ppa) and the distribution of pulmonary vascular resistance using the double-occlusion method. 3. ET-1 (10(-12)-10(-9) M) produced concentration-dependent pulmonary vasodilatation. ET-1 (10(-9) M) decreased Ppa from 24.5 +/- 3.1 to 17.0 +/- 3.0 cmH2O with a nadir occurring at 1 min, followed by a slow return to baseline over 60 min (time for half-recovery (t1/2R) of 17.2 min). The decrease in Ppa was the result of pulmonary precapillary vasodilatation. Endothelin-3 (ET-3) (10(-12) and 10(-11) M) also induced vasodilatation comparable to equimolar concentrations of ET-1, whereas the selective ETB receptor agonist IRL 1620 at equimolar concentrations caused a more protracted vasodilatation response. 4. Neither the cyclo-oxygenase inhibitor indomethacin (10(-5) M) nor the KATP+ (ATP-sensitive) potassium channel blocker glibenclamide (10(-5) M) significantly altered the baseline Ppa; moreover, neither inhibitor affected the ET-1-induced vasodilatation, indicating the lack of involvement of cyclo-oxygenase metabolites and KATP+ channel activity in the mediation of the pulmonary vasodilator response to ET-1. 5. Addition of 10(-5) M reduced haemoglobin, which antagonizes the action of nitric oxide (NO), increased Ppa over prehaemoglobin levels. Haemoglobin significantly decreased the duration (t1/2R, 3.8 +/- 0.7 min) of pulmonary vasodilatation to ET-1, but did not abolish the initial phase of the response. L-N-Monomethylarginine, an inhibitor of NO synthesis, either alone or in combination with haemoglobin, similarly reduced the duration of ET-1-induced pulmonary vasodilatation. 6. The ETA receptor antagonist [Dpr1-Asp15]-ET-1 (Dpr, diaminoproprionic acid) had no effect on pulmonary vasodilatation induced by ET-1, ET-3 or IRL 1620 (suc-(Glu9,Ala11,15)-ET-1(8-21)). This finding combined with the observed relative potencies of the peptides (IRL 1620 > ET-1 = ET-3) suggests that pulmonary vasodilatation was mediated by activation of the non-selective ETB receptor. 7. The results indicate that the sustained ET-1-induced pulmonary vasodilatation in neonates is probably mediated via ETB receptor activation and that it is critically dependent on NO.


Platinum(II) complexes with 5,7-disubstituted-1,2,4-triazolo [1,5-a]pyrimidines: spectroscopical characterization and cytotoxic activity in vitro.
Complexes of the types: cis-[PtI(2)(dptp)(2)] (1), cis-[PtI(2)(NH(3))(dptp)] (2), trans-[PtI(2)(dptp)(dmso)] (3) and trans-[PtI(2)(dbtp)(dmso)] (4), where dptp=5,7-diphenyl-1,2,4-triazolo[1,5-a]pyrimidine (dptp), dbtp=5,7-ditertbutyl-1,2,4-triazolo[1,5-a]pyrimidine have been synthesized and characterized by infrared and multinuclear magnetic resonance spectroscopic techniques ((1)H, (13)C, (15)N, (195)Pt). In (195)Pt NMR, the cis-diiodo complexes were observed between -2601 ppm and -3261 ppm, while the trans coordination compounds were found at higher field (ca. -4389 ppm). In all cases significant (15)N NMR shielding (92-95 ppm) were observed for N(3) atom indicating this nitrogen atom as a coordination site. The cis complexes have been assayed for antitumor activity in vitro against two human cell lines: A549 (non-small cell lung carcinoma) and T47D (breast cancer). The results indicate a moderate antiproliferative activity of (2) against human cancer lines.


Process Evaluation of a Digital Platform-Based Implementation Strategy Aimed at Work Stress Prevention in a Health Care Organization.
The objective was to evaluate the process and feasibility of a digital platform-based implementation strategy aimed at work stress prevention. The process evaluation was performed alongside a controlled trial within a health care organization, in the experimental group (N = 221). Mental models, context, and barriers and facilitators were measured. In addition, dose delivered, reach, and dose received were assessed. Employees reported relatively high readiness for change. Personnel shortage and a recent restructuring of the organization hindered use of the strategy. Low management support and high turnover stagnated strategy deployment. Dose delivered was 13/15, reach was 11/15, and dose received was 5/15. Strategy implementation was moderately successful, as sustained strategy use by the teams appeared to be a challenge. The strategy can be feasible with sufficient management support and resources.


Accumulation and biochemical effects of microcystin-LR on the Patagonian pejerrey (Odontesthes hatcheri) fed with the toxic cyanobacteria Microcystis aeruginosa.
We studied accumulation and biochemical effects of microcystin-LR (MCLR) in Odontesthes hatcheri after dietary administration of the cyanobacteria Microcystis aeruginosa (1.3 μg MCLR/g body mass, incorporated in standard fish food). After 12 h, MCLR content in liver did not differ between fish fed with crushed or intact cells, demonstrating O. hatcheri's capacity to digest cyanobacteria and absorb MCLR. In the second experiment, fish received toxic cells, non-toxic cells, or control food; MCLR accumulation was monitored for 48 h. Protein phosphatase 1 (PP1), catalase (CAT), glutathione-S-transferase (GST) activities, and lipid peroxidation (as MDA) were measured in liver and intestine. Methanol-extractable MCLR was determined by PP1 inhibition assay (PPIA); extractable and protein-bound MCLR were measured by Lemieux oxidation-gas chromatography/mass spectrometry (GC/MS). MCLR accumulated rapidly up to 22.9 and 9.4 μg MCLR/g in intestine and liver, respectively, followed by a decreasing tendency. Protein-bound MCLR represented 66 to ca. 100 % of total MCLR in both tissues. PP1 activity remained unchanged in intestine but was increased in liver of MCLR treated fish.CAT and GST activities and MDA content were significantly increased by MCLR only in liver. We conclude that O. hatcheri is able to digest cyanobacteria, accumulating MCLR mostly bound to proteins. Our data suggest that this freshwater fish can be adversely affected by cyanobacterial blooms. However, the rapid decrease of the detectable MCLR in both tissues could imply that sublethal toxin accumulation is rapidly reversed.


Tolerance to human gamma globulin in mice selected for high or low antibody responsiveness.
The injection of aggregate-free or tolerogenic human gamma globulin (t-HGG) induced tolerance in two lines of mice selected for high or low antibody responsiveness (high responder (H) or low responder (L) lines, respectively). In L mice, the tolerance state was very short and could no longer be obtained when the challenge injection was delayed for four weeks after tolerogenic treatment. At that time, an antibody response to t-HGG was observed in both lines; in H mice, however, this was followed by a partial tolerance state. These results and those previously obtained concerning tolerance induction to bovine serum albumin and rabbit gamma globulin in H and L mice suggest at least partial independent genetic control of antibody responsiveness and susceptibility to tolerance induction.


Studies on T cell subsets in chronic urticaria by monoclonal antibodies.
Peripheral blood T lymphocytes and T cell subsets were explored in 25 patients with chronic urticaria and 25 healthy controls. Thirteen of the patients had lymphocytic or leukocytoclasic vasculitis. An abnormal distribution between helper and suppressor T cell subsets with decreased percentage of helper (OKT4) T cells was found as compared to controls.


Muscular and hepatic pollution biomarkers in the fishes Phycis blennoides and Micromesistius poutassou and the crustacean Aristeus antennatus in the Blanes Submarine Canyon (NW Mediterranean).
Submarine canyons are regarded as a sink for pollutants. In order to determine if this theory applied to deep-sea species from an important fishing ground (the Blanes submarine canyon) located in the NW Mediterranean, we sampled the commercial fish Phycis blennoides and Micromesistius poutassou and the crustacean Aristeus antennatus. Specimens were sampled inside and outside (in the open continental slope) the submarine canyon; both are regarded as potentially affected by exposure to different anthropogenic chemicals. Several pollution biomarkers in muscle (activity of cholinesterases) and liver/hepatopancreas (catalase, glutathione S-transferases, carboxylesterases, ethoxyresorufin O-deethylase in fish or mixed function oxygenase (MFO)-related reductases in crustacean, and lipid peroxidation levels) were measured. Chemical analysis of the persistent organic pollutants, namely polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethanes (DDTs) and hexachlorocyclohexanes (HCHs) was also performed on the fish and crustacean muscle. Biomarker activities and levels were discussed in relation to pollutant exposure, habitat, and parameters including sex, size, and species. Biochemical responses and chemical analysis of PCBs evidenced interspecies differences as well as sex and size-related ones, mainly in A. antennatus. An indication of higher exposure to pollutants inside the canyon was observed, which was more clearly reflected in the fish than in the crustacean. However, further research is required to confirm this observation.


Pesticide leaching from two Swedish topsoils of contrasting texture amended with biochar.
The use of biochar as a soil amendment has recently increased because of its potential for long-term soil carbon sequestration and its potential for improving soil fertility. The objective of this study was to quantify the effects of biochar soil incorporation on pesticide adsorption and leaching for two Swedish topsoils, one clay soil and one loam soil. We used the non-reactive tracer bromide and the pesticides sulfosulfuron, isoproturon, imidacloprid, propyzamid and pyraclostrobin, substances with different mobility in soil. Adsorption was studied in batch experiments and leaching was studied in experiments using soil columns (20 cm high, 20 cm diameter) where 0.01 kg kg(-1) dw biochar powder originating from wheat residues had been mixed into the top 10 cm. After solute application the columns were exposed to simulated rain three times with a weekly interval and concentrations were measured in the effluent water. The biochar treatment resulted in significantly larger adsorption distribution coefficients (Kd) for the moderately mobile pesticides isoproturon and imidacloprid for the clay soil and for imidacloprid only for the loam soil. Relative leaching of the pesticides ranged from 0.0035% of the applied mass for pyraclostrobin (average Kd=360 cm3 g(-1)) to 5.9% for sulfosulfuron (average Kd=5.6 cm3 g(-1)). There were no significant effects of the biochar amendment on pesticide concentrations in column effluents for the loam soil. For the clay soil concentrations were significantly reduced for isoproturon, imidacloprid and propyzamid while they were significantly increased for the non-mobile fungicide pyraclostrobin suggesting that the transport was facilitated by material originating from the biochar amendment.


Evaluating the Accuracy of FUCCI Cell Cycle In Vivo Fluorescent Imaging to Assess Tumor Proliferation in Preclinical Oncology Models.
The primary goal of this study is to evaluate the accuracy of the fluorescence ubiquitination cell cycle indicator (FUCCI) system with fluorescence in vivo imaging compared to 3'-deoxy-3'-[18F]fluorothymidine ([18F]-FLT) positron emission tomography (PET)/computed tomography (CT) and biological validation through histology. Imaging with [18F]-FLT PET/CT can be used to noninvasively assess cancer cell proliferation and has been utilized in both preclinical and clinical studies. However, a cost-effective and straightforward method for in vivo, cell cycle targeted cancer drug screening is needed prior to moving towards translational imaging methods such as PET/CT. In this study, fluorescent MDA-MB-231-FUCCI tumor growth was monitored weekly with caliper measurements and fluorescent imaging. Seven weeks post-injection, [18F]-FLT PET/CT was performed with a preclinical PET/CT, and tumors samples were harvested for histological analysis. RFP fluorescent signal significantly correlated with tumor volume (r = 0.8153, p < 0.0001). Cell proliferation measured by GFP fluorescent imaging was correlated with tumor growth rate (r = 0.6497, p < 0.001). Also, GFP+ cells and [18F]-FLT regions of high uptake were both spatially located in the tumor borders, indicating that the FUCCI-IVIS method may provide an accurate assessment of tumor heterogeneity of cell proliferation. The quantification of total GFP signal was correlated with the sum of tumor [18F]-FLT standard uptake value (SUV) (r = 0.5361, p = 0.0724). Finally, histological analysis confirmed viable cells in the tumor and the correlation of GFP + and Ki67 + cells (r = 0.6368, p = 0.0477). Fluorescent imaging of the cell cycle provides a noninvasive accurate depiction of tumor progression and response to therapy, which may benefit in vivo testing of novel cancer therapeutics that target the cell cycle.


[Basal cell carcinoma among the cases of Dermatology Clinic I of the Silesian Medical Academy in Katowice].
The authors present their own experiences in the treatment of 24 patients with the nevoid basal-cell carcinoma syndrome. All the patients underwent surgical treatment, in 12 of them some of the tumours were treated with X-rays. Two patients were found to have 32 and 26 nevoid basal-cell carcinoma respectively. In none of the 24 patients recurrence was noted. However in many of them the new foci of carcinoma required systemic treatment.


Molecular epidemiology of carbapenem-resistant Acinetobacter baumannii isolated from three major hospitals in Jordan.
In the last decade, incidences of carbapenem-resistant Acinetobacter baumannii have been increasingly reported worldwide. Consequently, A. baumannii was included in the World Health Organization's new list of critical pathogens, for which new drugs are desperately needed. The objective of this research was to study the molecular epidemiology and antimicrobial susceptibility of clinical carbapenem-resistant A. baumannii isolated from Jordanian hospitals. A total of 78 A. baumannii and 8 Acinetobacter spp. isolates were collected from three major hospitals in Jordan during 2018. Disc diffusion and microdilution methods were used to test their susceptibility against 19 antimicrobial agents. Multilocus sequence typing (MLST) was performed using the Pasteur scheme, followed by eBURST analysis for all isolates. PCR was used to detect β-lactam resistance genes, blaOXA-23-like , blaOXA-51-like , and blaNDM-1 . Of the 86 tested isolates, 78 (90.6%) exhibited resistance to carbapenems, whereas no resistance was recorded to tigecycline or polymyxins. Based on the resistance profiles, 10.4% and 84.8% of isolates were classified into multidrug resistant (MDR) or extensively drug resistant (XDR), respectively. The most prevalent carbapenems resistance genes amongst isolates were blaOXA-51-Like (89.5%), followed by blaOXA-23-Like (88.3%) and blaNDM-1 (10.4%). MLST revealed the presence of 19 sequence types (STs), belonging to eight different international complexes. The most commonly detected clonal complex (CC) was CC2, representing 64% of all typed isolates. This is the first study to report the clonal diversity of A. baumannii isolates in Jordan. A high incidence of carbapenem resistance was detected in the isolates investigated. In addition, our findings provided evidence for the widespread of blaOXA-23-like harbouring carbapenem-resistant A. baumannii and belonging to CC2. The number of XDR isolates identified in this study is alarming. Thus, periodic surveillance and molecular epidemiological studies of resistance factors are important to improve treatment outcomes and prevent the spread of A. baumannii infections.


Bone Marrow Mesenchymal Stem Cell-Based Engineered Cartilage Ameliorates Polyglycolic Acid/Polylactic Acid Scaffold-Induced Inflammation Through M2 Polarization of Macrophages in a Pig Model.
: The regeneration of tissue-engineered cartilage in an immunocompetent environment usually fails due to severe inflammation induced by the scaffold and their degradation products. In the present study, we compared the tissue remodeling and the inflammatory responses of engineered cartilage constructed with bone marrow mesenchymal stem cells (BMSCs), chondrocytes, or both and scaffold group in pigs. The cartilage-forming capacity of the constructs in vitro and in vivo was evaluated by histological, biochemical, and biomechanical analyses, and the inflammatory response was investigated by quantitative analysis of foreign body giant cells and macrophages. Our data revealed that BMSC-based engineered cartilage suppressed in vivo inflammation through the alteration of macrophage phenotype, resulting in better tissue survival compared with those regenerated with chondrocytes alone or in combination with BMSCs. To further confirm the macrophage phenotype, an in vitro coculture system established by engineered cartilage and macrophages was studied using immunofluorescence, enzyme-linked immunosorbent assay, and gene expression analysis. The results demonstrated that BMSC-based engineered cartilage promoted M2 polarization of macrophages with anti-inflammatory phenotypes including the upregulation of CD206, increased IL-10 synthesis, decreased IL-1β secretion, and alterations in gene expression indicative of M1 to M2 transition. It was suggested that BMSC-seeded constructs have the potential to ameliorate scaffold-induced inflammation and improve cartilaginous tissue regeneration through M2 polarization of macrophages. Finding a strategy that can prevent scaffold-induced inflammation is of utmost importance for the regeneration of tissue-engineered cartilage in an immunocompetent environment. This study demonstrated that bone marrow mesenchymal stem cell (BMSC)-based engineered cartilage could suppress inflammation by increasing M2 polarization of macrophages, resulting in better tissue survival in a pig model. Additionally, the effect of BMSC-based cartilage on the phenotype conversion of macrophages was further studied through an in vitro coculture system. This study could provide further support for the regeneration of cartilage engineering in immunocompetent animal models and provide new insight into the interaction of tissue-engineered cartilage and macrophages.


Molecular characterization of frog chromogranin B reveals conservation of selective sequences encoding potential novel regulatory peptides.
Chromogranin B (CgB) is a member of the granin family of neuroendocrine secretory proteins, which has been proposed to play a role in secretory granule biogenesis and as a precursor to bioactive peptides. The cloning of CgB in a phylogenetically distant vertebrate, the frog Rana ridibunda, reveals a modest overall homology (35-40%) with mammalian CgB. However, the sequences of the N- and C-terminal regions are more highly conserved (57-65% amino acid identity) and may give rise to novel regulatory peptides. In frog, intense expression of CgB mRNA was observed in particular structures of the brain and in the distal lobe of the pituitary.


Increased cytotoxicity of ricin in a putative Golgi-defective mutant of Chinese hamster ovary cell.
We have studied the cytotoxicity of ricin in a monensin-resistant mutant (MonR-31) of Chinese hamster ovary (CHO) cell line which is presumably altered in Golgi functions/structures. The cytotoxicity of ricin was increased in MonR-31 mutant cells compared with that in its parental CHO cells. In wild-type CHO cells, the cytotoxicity of ricin was enhanced by HN4Cl, bafilomycin A1, or nigericin. The enhancement of ricin cytotoxicity by these compounds was greatly reduced in MonR-31 mutant cells. Brefeldin A (BFA), which disrupts the structure of the Golgi apparatus, inhibits the cytotoxicity of ricin in both CHO and MonR-31 cells. We have also examined the effects of glycosylation inhibitors and the removal of high mannose oligosaccharide chains in ricin on the ricin hypersensitivity in MonR-31 cells. The hypersensitivity of MonR-31 cells to ricin is apparently not due to any difference in glycosylation between CHO and MonR-31 cells or in the processing of oligosaccharides on ricin by the target cells. Nigericin at low concentration (10 nM), which has no effect on the cytotoxicity of diphtheria toxin, enhances the ricin cytotoxicity, but inhibits the modeccin cytotoxicity. Our results suggest that important step(s) in the intoxication process of CHO cells by ricin and modeccin take place in the Golgi region.


Successful pregnancy in a liver transplant recipient following controlled ovarian hyperstimulation and intracytoplasmic sperm injection.
As the demand for assisted reproductive technologies (ART) increases, infertility treatments are increasingly being sought by patients with serious health problems that were once regarded as leading to infertility. Moreover, since pregnancy may worsen their underlying health conditions, these women have been advised to remain infertile. We describe here a liver transplant recipient who underwent ART for male factor infertility. Assisted fertilization was achieved using intracytoplasmic sperm injection, and she conceived after embryo transfer. Her pregnancy follow-up was unremarkable until she delivered a healthy boy pretermly at 31 and half weeks.


[Orbital subperiotal hematoma associated with extradural hematoma: one case report].
Subperiotal hematoma of the orbit associated with extradural hematoma has been rarely reported in the literature. We report a case of an 8-year-old girl who presented after traumatism, a subperiotal hematoma of the orbit with two extradural hematomas. During the first clinical examination realized while the patient was unconscious, left exophthalmia was found, but after conscious improvement, we discovered ophtalmoplegia and a grade I papilla edema. The CT scan showed two extradural hematomas and a left intraorbital hematoma. The intraorbital hematoma was drained by surgical way but the extradural hematomas were not surgical. The outcome has been marked by the neurological improvement and the oculomotricity recovering.


Longitudinal research on the bidirectional association between depression and arthritis.
Studies evaluating the mutual relation between depression and arthritis have been limited and yielded inconsistent results. The aim of this study was to investigate the bidirectional relationship between depression and arthritis in a middle-aged and elderly Chinese population. Participants ≥ 45 years of age were included from the China Health and Retirement Longitudinal Study (CHARLS). In stage I, we assessed the association of baseline depression with follow-up arthritis. In stage II, we examined whether the onset of arthritis predicted future depression. Logistic regression analyses were conducted to estimate the odds ratios (ORs) and confidence intervals (CIs) in stage I and stage II, respectively. In stage I, 24.3% (679/2794) of the depression group and 15.4% (1000/6482) of the non-depression group developed new arthritis cases. Compared with non-depression individuals, the risk of developing arthritis in depression patients was significantly higher (OR: 1.56, 95% CI 1.37-1.79). In stage II, 39.7% (973/2453) subjects in the arthritis group and 26.7% (1667/6236) subjects in the non-arthritis group developed depressive symptoms. The adjusted OR (95% CI) for depression in the arthritis group was 1.64 (1.45-1.86) times higher than that in the non-arthritis group. In the subgroup analyses according to sex, age, household income, residence, body mass index, smoking and drinking, all sub-groups yielded consistent associations. The onset of depression increased the risk of incident arthritis; in addition, baseline arthritis predicted future depression in middle-aged and elderly Chinese adults.


Blockade of FGF2/FGFR2 partially overcomes bone marrow mesenchymal stromal cells mediated progression of T-cell acute lymphoblastic leukaemia.
The development of acute lymphoblastic leuakemia (ALL) is partly attributed to the effects of bone marrow (BM) microenvironment, especially mesenchymal stromal cells (MSCs), which interact bilaterally with leukaemia cells, leading to ALL progression. In order to find MSCs-based microenvironment targeted therapeutic strategies, Notch1-induced T-cell ALL (T-ALL) mice models were used and dynamic alterations of BM-MSCs with increased cell viability during T-ALL development was observed. In T-ALL mice derived stroma-based condition, leukaemia cells showed significantly elevated growth capacity indicating that MSCs participated in leukaemic niche formation. RNA sequence results revealed that T-ALL derived MSCs secreted fibroblast growth factor 2 (FGF2), which combined with fibroblast growth factor receptor 2 (FGFR2) on leukaemia cells, resulting in activation of PI3K/AKT/mTOR signalling pathway in leukaemia cells. In vitro blocking the interaction between FGF2 and FGFR2 with BGJ398 (infigratinib), a FGFR1-3 kinase inhibitor, or knockdown FGF2 in MSCs by interference caused deactivation of PI3K/AKT/mTOR pathway and dysregulations of genes associated with cell cycle and apoptosis in ALL cells, leading to decrease of leukaemia cells. In mouse model received BGJ398, overall survival was extended and dissemination of leukaemia cells in BM, spleen, liver and peripheral blood was decreased. After subcutaneous injection of primary human T-ALL cells with MSCs, tumour growth was suppressed when FGF2/FGFR2 was interrupted. Thus, inhibition of FGF2/FGFR2 interaction appears to be a valid strategy to overcome BM-MSCs mediated progression of T-ALL, and BGJ398 could indeed improve outcomes in T-ALL, which provide theoretical basis of BGJ398 as a BM microenvironment based therapeutic strategy to control disease progression.


The maize zein gene zE19 contains two distinct promoters which are independently activated in endosperm and anthers of transgenic Petunia plants.
The activity, tissue specificity and temporal expression of the tandem promoter region preceding a maize zein gene (zE19, encoding a 19 kDa zein protein) were tested in transgenic Petunia plants. To simplify the analysis, the tandem promoter as well as each of the two separate promoter regions were fused to the beta-glucuronidase (GUS) reporter gene. All of the three constructs directed the synthesis of GUS in the endosperm of transformed seeds indicating that both separate promoters are independently activated and show the same tissue and cell type specificity observed for zein genes in maize. The kinetics of accumulation and the localization of GUS activity are not coordinated with those of Petunia endogenous seed storage proteins during the development of transformed seeds. Unexpectedly, we detected high levels of GUS activity in anthers of transformed Petunia plants for all three constructs. This appears to reflect the expression pattern of zein genes in maize, since we detect zein transcripts in anthers. Finally, we discuss the possible origin and function of the tandem promoter arrangement on the basis of these data.


[Chondrosarcoma of the larynx: review of the Western literature and presentation of a new case].
We present the case of a 70 year old man with cricoid chondrosarcoma treated with total laryngectomy. This report reviews the occidental literature, with nine cases reported by national authors in the las 25 years. We have recollected 104 laryngeal chondrosarcoma, in all. Clinical and diagnostical data, location, and treatment were analysed and compared with our case.


A cross-sectional study of intestinal parasitoses in dogs and children of the periurban area of La Plata (Buenos Aires, Argentina): Zoonotic importance and implications in public health.
A cross-sectional study was performed between school term dates 2014 and 2015 to diagnose intestinal parasites in dogs and children living with them. The socio-environmental characteristics and hygiene practices of the children were also evaluated in terms of risk factors for parasitic infection of periurban neighbourhoods of La Plata (Buenos Aires, Argentina). Serial coproparasitological samples of 78 dogs and 211 children were analysed by means of concentration and flotation techniques. Socio-environmental variables and hygiene practices of children were evaluated through semi-structured questionnaires which were answered by every family. The study showed that 82.1% of dogs were parasitized. The specific richness was of 11 species; Ancylostoma caninum (69.2%), Uncinaria stenocephala (41.0%), Trichuris vulpis (28.2%) and Toxocara canis (21.8%) were the most prevalent. The study also revealed that 67.8% of children were positive. Also, 11 species were identified and the most prevalent were Blastocystis sp. (36.0%), Enterobius vermicularis (27.5%) and Giardia lamblia (21.3%). The risk for parasitosis was higher in 6-year-old children and older (OR = 1.9, 95% IC: 1.0-3.7) and in those who did not wash their hands or did it occasionally after playing with their pets (OR = 2.8, 95% IC: 1.4-5.5). Blastocystis sp. and Entamoeba coli infection risks were greater in children whose parents had a basic level of education (OR = 3.4, 95% IC: 1.3-8.7 and OR = 3.6, 95% IC: 0.8-15.9, respectively). In addition, the risk of infection for E. coli was higher in children who lived in floodable houses (OR = 4.4, 95% IC: 0.9-16.6). Likewise, the risk of infection for E. vermicularis was greater in children with onychophagia (OR = 1.6, 95% IC: 0.7-3.7) and in 6 year olds and older whose parents completed only primary studies (OR = 3.6, 95% IC: 1.4-9.1). The results obtained show the existence of a worrying epidemiological scenario that stresses the importance of zoonotic parasitosis as a serious problem of public health.


A world key to the genera of Elcanidae (Insecta, Orthoptera), with a Jurassic new genus and species of Archelcaninae from China.
A new fossil genus and species is described from the Middle Jurassic of China. The type of Sinoelcana minuta gen. et sp. nov. has body and legs preserved. It is distinguished from all other elcanids by the unique combination of wing venation and stout ovipositor. The sickle-shaped ovipositor suggests that the new species had a preference for oviposition on plant material. A world key to the genera of Elcanidae is provided based on the wing venation.


Robotic-assisted proctectomy for inflammatory bowel disease: a case-matched comparison of laparoscopic and robotic technique.
The objective of this study was to compare short-term outcomes of robotic and laparoscopic proctectomy in patients with inflammatory bowel disease (IBD). This is an IRB-approved case-matched review. Seventeen robotic proctectomies (RP), 10 with ileal pouch anal anastomosis (IPAA) and 7 completion (CP), were matched to laparoscopic proctectomies (LP). Short-term and functional outcomes were compared between LP and RP. In CP cohort, operative times were longer in the RP group (351 RP vs 238 LP min, p = 0.03), mean robotic time 90 min. Estimated blood loss (EBL) was similar between RP-CP and LP-CP groups (p = 0.18). Return of bowel function (RBF) was slower in RP-CP group (3.0 vs 1.7 days, p = 0.04), and length of stay (LOS) was longer (6.4 vs 4.1 days, p = 0.02). In the IPAA group, there were no differences between operative times (p = 0.14), robotic time 86 min; EBL (p = 0.15), and postoperative complications. Return of bowel function (3.6 vs 2.6 days, p = 0.3) and LOS (8.5 vs 6.1 days, p = 0.17) were similar between RP and LP. Bowel and sexual function were equivalent between LP and RP-IPAA groups. Robotic proctectomy is a safe and effective technique for patients with IBD. It is comparable to LP with regard to perioperative outcomes, complications, and short-term functional results.


Microbial high cell density fermentations in a stirred single-use bioreactor.
: Microbial fermentations are of major importance in the field of biotechnology. The range of applications is rather extensive, for example, the production of vaccines, recombinant proteins, and plasmids. During the past decades single-use bioreactors have become widely accepted in the biopharmaceutical industry. This acceptance is due to the several advantages these bioreactors offer, such as reduced operational and investment costs. Although this technology is attractive for microbial applications, its usage is rarely found. The main limitations are a relatively low oxygen transfer rate and cooling capacity. The aim of this study was to examine a stirred single-use bioreactor for its microbial suitability. Therefore, the important process engineering parameters volumetric mass transfer coefficient (k L a), mixing time, and the heat transfer coefficient were determined. Based on the k L a characteristics a mathematical model was established that was used with the other process engineering parameters to create a control space. For a further verification of the control space for microbial suitability, Escherichia coli and Pichia pastoris high cell density fermentations were carried out. The achieved cell density for the E. coli fermentation was OD600 = 175 (DCW = 60.8 g/L). For the P. pastoris cultivation a wet cell weight of 381 g/L was reached. The achieved cell densities were comparable to fermentations in stainless steel bioreactors. Furthermore, the expression of recombinant proteins with titers up to 9 g/L was guaranteed.


Molecular determination of Tritrichomonas spp. in aborted bovine foetuses in Eastern Anatolian Region of Turkey.
Tritrichomonas foetus is the causative agent of venereal trichomonosis in cattle causing infertility, pyometra and abortions. The objectives of this study were to determine the positivity rate of Tritrichomonas spp. in abomasal content of aborted foetuses from Eastern Anatolian Region of Turkey, using staining, culture and PCR methods and to present the isolates found in the region. A total of 246 abomasal content of aborted foetuses were tested and 14 of 246 (5.7%) were Tritrichomonas spp. positive only by polymerase chain reaction (PCR). Positivity was not attained by staining or culture method. Four of the positive samples in PCR were confirmed to be T. foetus by sequencing of the amplified 5.8S rRNA gene and flanking ITS regions. Nucleotide sequences of TR-Erzurum T. foetus isolates have been entered into the GenBank sequence database under accession numbers KC236423 through KC236426. This preliminary study suggests that future studies are needed on the systematic relationships and epidemiology of T. foetus isolates in the region.


An analysis of neuronal elements within the median nucleus of the raphe that mediate lesion-induced increases in locomotor activity.
Electrolytic lesions of the median nucleus of the raphe (MR) are known to result in large increases in motor activity. The present studies were concerned with identifying the neuronal elements within or near the MR (e.g. fibers of passage, serotonergic or non-serotonergic cells) which, when destroyed, lead to these increases in ambulation. Groups of rats were given either an electrolytic MR lesion or were injected locally with the serotonin neurotoxin 5,7-dihydroxytryptamine (5,7-DHT) or the excitotoxin ibotenic acid, and their subsequent locomotor activities were compared in the open field and in photocell cages. In a 5 min open field test, rats with either electrolytic or ibotenate lesions of the MR were more active compared to all other groups, although rats in the former group were also more active than those in the latter. In a longer activity test conducted in photocell cages, rats with electrolytic lesions were more active than all other groups during the first 20 min period, after which their activity did not differ from the ibotenate group; however, both of these groups were hyperactive compared to all others. Both electrolytic and ibotenate groups showed exaggerated hyperactivity in response to D-amphetamine. Lesions produced by 5,7-DHT failed to significantly increase either spontaneous or D-amphetamine-induced locomotor activity. When the results from all of the activity measures are considered, it appears that damage to at least two different neuronal populations may be responsible for the hyperactivity observed in MR lesioned rats. However, damage to the serotonergic system does not appear to contribute to these locomotor effects.


Anatomical Landmark Technique for Femoral Tunnel Placement of Lateral Extra-Articular Tenodesis.
The anterolateral ligament is a crucial part of the anterolateral complex of the knee, providing rotator stability to the knee and being a primary restraint to tibial internal rotation. Lateral extra-articular tenodesis added to anterior cruciate ligament reconstruction can reduce pivot shift without sacrificing the range of motion or increasing the risk of osteoarthritis. A 7- to 8-cm longitudinal skin incision is made and a 9.5- to 10-cm × 1- cm wide iliotibial band graft is dissected, leaving the distal attachment intact. The free end is whip stitched. One of the most important steps during the procedure is the identification of the site of attachment of the iliotibial band graft. The leash of vessels, fat pad, lateral supracondylar ridge, and fibular collateral ligament serve as important landmarks. The tunnel is drilled from the lateral femoral cortex with a guide pin and reamer pointing 20 to 30° anteriorly and proximally while the arthroscope visualizes the femoral anterior cruciate ligament tunnel. The graft is routed under the fibular collateral ligament. The graft is fixed with a bioscrew while the knee is kept in 30° flexion and the tibia is kept in neutral rotation. We believe that lateral extra-articular tenodesis gives the anterior cruciate ligament graft a good chance for faster healing along with addressing anterolateral rotatory instability. Choosing a correct fixation point is very important to restore normal knee biomechanics.


Compression therapy for leg disorders: Critique 2.
This paper reviews compression therapy of the lower extremities and lays down guidelines for the levels of compression that are required to counteract different degrees of increased tissue pressure.


Dolutegravir Population Pharmacokinetics in a Real-Life Cohort of People Living With HIV Infection: A Covariate Analysis.
There are growing concerns about dolutegravir (DTG)-related neuropsychiatric adverse events and about differences in the characteristics of people living with HIV infection (PLWH) potentially associated with higher risks of said side effects. Several studies have shown that DTG was stopped more frequently among women, older PLWH, and PLWH who initiated abacavir (ABC) at the same time. This study aimed to clarify the factors affecting the pharmacokinetics (PKs) of DTG in a real-life cohort of PLWH using a population PK approach. The model-building strategy was based on a previously published model developed from premarketing trials (1-compartment model with first-order absorption and a lag time). Sparse therapeutic drug monitoring data were obtained from a real-life cohort of 279 PLWH, and population PK analysis was performed using Monolix software. A stepwise covariate model-building strategy was used to evaluate any relevant effects of age, body weight, gender, total bilirubin, smoking status, formulations of DTG, morning versus evening dosing, backbone therapy, and other comedications including CYP/UGT inducers/inhibitors. For a typical 70-kg PLWH, the apparent clearance (CL/F) and apparent volume of distribution (V/F) were 0.748 L/h and 14.6 L, respectively. Of the demographic factors evaluated, body weight was a significant covariate for CL/F and for V/F. Smokers had a 17% higher CL/F relative to nonsmokers. Both strong enzyme inhibitors (eg, atazanavir) and inducers (eg, rifampicin) had marked effects on DTG exposure, with potential clinical implications. Ritonavir-boosted darunavir was found to moderately increase clearance of DTG by 23%. No significant effect of ABC-based backbone therapy was observed on the PK parameters of DTG. Our results did not support the hypothesis that ABC, by competing with the DTG metabolic pathway, may significantly increase DTG exposure leading to potential drug toxicity.


Effects of prolactin during preincubation of mouse spermatozoa on fertilizing capacity in vitro.
In the present study, some beneficial effects of mouse prolactin (PRL) on spermatozoa were demonstrated in mice. The motility rate of spermatozoa is well maintained during incubation in modified Krebs-Ringer bicarbonate solution (mKRB) containing PRL (10 or 100 ng/ml) for up to 120 min. When spermatozoa that had been preincubated in the same PRL-containing mKRB were incubated with oocytes in mKRB, significantly more spermatozoa attached to the zona pellucida of each oocyte. When spermatozoa that had been preincubated in mKRB with PRL (50 and 100 ng/ml) for only 15 or 30 min were used for in vitro fertilization (IVF), significantly higher fertilization rates were yielded by those spermatozoa than by control spermatozoa preincubated without PRL for the same periods. The prolongation of the preincubation period did not result in increased fertilization rates. Thus, PRL demonstrated biological effects on spermatozoa by shortening the optimal preincubation period for spermatozoa to acquire capacitation and by maintaining their motility and ability of the attachment to the oocyte during IVF. The results are relevant to clinical application of PRL for infertile patients with oligozoospermia or asthenozoospermia.


Inhibition of Dengue virus and West Nile virus proteases by click chemistry-derived benz[d]isothiazol-3(2H)-one derivatives.
Two click chemistry-derived focused libraries based on the benz[d]isothiazol-3(2H)-one scaffold were synthesized and screened against Dengue virus and West Nile virus NS2B-NS3 proteases. Several compounds (4l, 7j-n) displayed noteworthy inhibitory activity toward Dengue virus NS2B-NS3 protease in the absence and presence of added detergent. These compounds could potentially serve as a launching pad for a hit-to-lead optimization campaign.


Culturally safe, trauma-informed approach to cognitive impairment and dementia in older Aboriginal and Torres Strait Islander people.
The population of older (age ≥50 years) Aboriginal and Torres Strait Islander people is increasing, and the rate of dementia in this group is three to fivefold higher than in the general population. A high prevalence of risk factors across the life course likely contributes to these rates. This paper highlights practical ways to apply best practice principles when addressing the detection of cognitive impairment and dementia in older Aboriginal and Torres Strait Islander people and its management in the context of a person's family, carers and community. Recognising culture, the collective nature of family and community decision making for a person, the ways to uphold respect for Elders and a trauma-informed approach are fundamental to the care of older Aboriginal and Torres Strait Islander people with cognitive impairment and dementia.


Variations of HRV and skin conductance reveal the influence of CV4 and Rib Raising techniques on autonomic balance: A randomized controlled clinical trial.
The aim of the present study was to evaluate whether Fourth Ventricle Compression Technique (CV4) and Rib Raising (RR) osteopathic techniques influence Autonomic Nervous System (ANS) activity, as measured by Heart Rate Variability (HRV) and Skin Conductance (SC). A randomized-controlled clinical trial has been performed from June 2010 to January 2011. 32 healthy adults (33.9 ± 14 years, and 72% female) were selected. Subjects were randomized in three groups: CV4 group, RR group and Placebo group. Each subject of each group underwent respective technique only once. HRV and SC were continuously recorded during the session. All data analysis was performed using SPSS statistical software (version 21.0) and the significance level was considered at p ≤ 0.05. RR technique intra-group subject analysis showed a significant decrease in LF/HF ratio (F = 25.18; p < 0.001), a statistically significant decrease of LF (F = 27.09; p < 0.001), and increase of HF, both at the end of treatment (F = 27.09; p < 0.001). CV4 technique intra-group subject analysis showed a significant reduction of the LF/HF ratio (F = 81.15; p < 0.001), a significant decrease in LF (F = 38.29; p < 0.001) and a significant increase of HF, at the end of treatment (F = 38.28; p < 0.001). Between-group analysis showed only a significant difference in LF/HF ratio between CV4 and PL groups (F = 3.44; p = 0.042). Our results suggest that the application of CV4 and RR techniques favors a shift in the autonomic balance towards a parasympathetic predominant state. Thus, their use within OMT protocol, as holistic approach, might improve the outcome of the treatment of clinical neuro-vegetative conditions.


Dermatologic Manifestations of Neurofibromatosis Type 1 and Emerging Treatments.
Neurofibromatosis type 1 (NF1) is an autosomal dominant tumor predisposition syndrome that increases one's risk for both benign and malignant tumors. NF1 affects every organ in the body, but the most distinctive symptoms that are often the most bothersome to patients are the cutaneous manifestations, which can be unsightly, cause pain or pruritus, and have limited therapeutic options. In an effort to increase awareness of lesser-known dermatologic associations and to promote multidisciplinary care, we conducted a narrative review to shed light on dermatologic associations of NF1 as well as emerging treatment options. Topics covered include cutaneous neurofibromas, plexiform neurofibromas, diffuse neurofibromas, distinct nodular lesions, malignant peripheral nerve sheath tumors, glomus tumors, juvenile xanthogranulomas, skin cancer, and cutaneous T-cell lymphoma.


Risk of endometrial polyps in women with endometriosis: a meta-analysis.
Endometrial polyps (EP) and endometriosis are both estrogen-dependent overgrowths of the endometrium. Several studies have shown a higher frequency of EP in endometriosis patients when compared with women without endometriosis. Therefore, we performed a meta-analysis to investigate the risk of EP in women with endometriosis. This meta-analysis searched for articles published between 1964 and 2014 in PubMed, Embase, and Cochrane Library, as well as in Chinese databases, including CNKI, VIP and Wanfang, regarding the association between endometriosis and EP. Nine cohort studies and one case-control study including 2896 women were included in this meta-analysis. The EP risk was evaluated using relative risk (RR) with a 95% confidence interval (CI). Heterogeneity, small study effect and publication bias were assessed using Higgins I(2), sensitivity analysis and funnel plots, respectively. The risk of EP increased in women with endometriosis compared with those without endometriosis (the pooled RR, 2.81; 95% CI, 2.48-3.18). No significant heterogeneity, small study effect or publication bias was found. The risk of EP slightly increased in women with endometriosis at stages 2-4 compared with those at stage 1 (Pooled effect size: stage 2 versus stage 1, RR, 1.22, 95% CI, 1.04 - 1.42; stage 3 versus stage 1, RR, 1.23, 95% CI, 1.06-1.42; stage 4 versus stage 1, RR, 1.29, 95% CI, 1.11-1.51; stages 2-4 versus stage 1, RR, 1.24, 95% CI, 1.10-1.40); however, no significantly different risk of EP in women with endometriosis existed between the other stages. The results suggest that it is important to identify whether patients with endometriosis also have EP and then remove any coexisting EP via hysteroscopy, especially for infertile patients. This process will be clinically helpful to treat endometriosis-related infertility in patients with endometriosis, especially for those with endometriosis that is more serious than stage 1.


Mathematical Models to Shed Light on Amyloid-Beta and Tau Protein Dependent Pathologies in Alzheimer's Disease.
The number of patients suffering from dementia due to Alzheimer's disease (AD) is constantly rising worldwide. This has accordingly resulted in huge burdens on the health systems and involved families. Lack of profound understanding of neural networking in normal brain and their interruption in AD makes the treatment of this neurodegenerative multifaceted disease a challenging issue. In recent years, mathematical and computational methods have paved the way towards a better understanding of the brain functional connectivity. Thus, much attention has been paid to this matter from both basic science researchers and clinicians with an interdisciplinary approach to determine what is not functioning properly in AD patients and how this malfunctioning can be addressed. In this review, a number of AD-related articles and well-studied pathophysiologic topics (e.g., amyloid-beta, neurofibrillary tangles, Ca2+ dysregulation, and synaptic plasticity alterations) has been literally surveyed from a computational and systems biology point of view. The neural networks were discussed from biological and mathematical point of views and their alterations in recent findings were further highlighted. Application of the graph theoretical analysis in the brain imaging was reviewed, depicting the relations between brain structure and function, without diving into mathematical details. Moreover, differential rate equations were briefly articulated, emphasizing the potential use of these equations in simplifying complex processes in relevance to pathologies of AD. Comprehensive insights were given into the AD progression from neural networks perspective, which may lead us towards potential strategies for early diagnosis and effective treatment of AD.


Developmental changes of neuron-specific enolase in human brain: an immunohistochemical study.
Developmental changes of neurons containing neuron-specific enolase (NSE) in human brain were studied in various areas of the central nervous system by immunohistochemistry with the peroxidase-antiperoxidase (PAP) method. In the brain stem, Purkinje cells, dentate nucleus, globus pallidus and thalamus, the number of NSE-positive neurons increased from an early period in gestation. However, in the pontine nucleus and putamen, it gradually increased along with decreasing cellularity later in gestation and in the infantile period. In the cerebral cortex, NSE-positive neurons developed as late as in the putamen and their cellularity increased earlier in the 5th layer than in the 3rd layer. Developmental changes of NSE-positive neurons parallel phylogenesis. The appearance of NSE-positive neurons can be a marker of neuronal maturation.


[The use of the pulse radiolysis method in the study of rapid processes in suspensions of Escherichia coli].
It was proved that the method of pulse radiolysis can be used to study the processes rapidly occurring in a microorganism suspension. It was shown that optically active particles formed after irradiation in an E. coli cell were involved into the first-(pseudo-first)-order reaction the rate constant being approximately 30 s-1. The presence of oxygen did not influence the kinetics of disappearance of these particles.


Effect of a very low negative dietary cation-anion difference (DCAD) diet on plasma and urine metabolomics of prepartum Holstein cows.
The objectives of this cross-sectional, nonintervention, observational study were to compare urine and blood parameters between cows consuming a positive dietary cation-anion difference (DCAD) diet [early dry cows, DCAD + 250 mEq/kg of dry matter (DM), n = 15] with the same cows consuming a negative DCAD diet (-220 mEq/kg of DM) 10 d after moving them from the early dry to the prepartum group. The most remarkable finding was that cows consuming the anionic diet had very low urine pH and very low base excess in blood, suggestive of uncompensated metabolic acidosis. Importantly, the metabolomics data revealed that only urine concentrations of essential and aromatic amino acids were decreased, and that concentrations of total nonessential amino acids and glucogenic amino acids were increased in plasma and reciprocally decreased in urine, suggesting that the cows fed anionic salts were attempting to meet a high glucose demand by mobilizing gluconeogenic amino acid reserves. Notably, the dietary anionic salts exerted marked effects on glycerophospholipids, with a reduction in most phosphatidylcholine containing diacyl (PC aa) and acyl-alkyl (PC ae) moieties in plasma and urine. Further characterization of these metabolomic profiles may lead to the development of novel biomarkers to identify cows susceptible to metabolic acidosis and other metabolic diseases.