Colony stimulating factor-1-induced osteoclast spreading depends on substrate and requires the vitronectin receptor and the c-src proto-oncogene.
The colony stimulating factor 1 (CSF-1) regulates osteoclastogenesis and bone resorption. Mutations in the CSF-1 gene cause an osteopetrosis characterized by the absence of osteoclasts. Mature osteoclasts respond to CSF-1 with inhibition of bone resorption and an increment of cell spreading. Herein we demonstrate that CSF-1-induced osteoclast spreading depends on the substrate the osteoclast interacts with and requires integrity of the vitronectin receptor and of the c-src proto-oncogene. Rabbit osteoclasts were allowed to attach to glass, serum, osteopontin, and bone substrates, and were treated with 10 ng/ml human recombinant CSF-1 for 4 h. In osteoclasts plated on glass, the cytokine induced 70% inhibition of bone resorption and 1.8-fold stimulation of cell spreading, without changes in podosome expression and microfilament array. In contrast, CSF-1 induced a 2.5-fold increase of osteoclasts showing filopodia, and a 9.5-fold increase of osteoclasts presenting lamellipodia, indicating that membrane motility was required for cell spreading. Osteoclasts plated on serum substrates showed a 50% reduction of spontaneous spreading. However, in this circumstance, CSF-1 still stimulated an increase of osteoclast area. In osteoclasts cultured on osteopontin substrate or on bone slices, an inhibition of CSF-1-induced osteoclast spreading was observed. To establish involvement of the vitronectin receptor and c-src proto-oncogene, cells were treated with the alpha vbeta3 integrin neutralizing antibody, LM609, or c-src antisense oligonucleotides, which reduced CSF-1-induced osteoclast spreading by 57% and 60%, respectively. The results demonstrate that CSF-1-induced osteoclast spreading requires both the vitronectin receptor and the c-src proto-oncogene and that this action is modulated by the adhesion substrata.


[3-D X-ray contrast rotational angiography in vascular pathology diagnostics].
The experience of 3-D X-ray contrast rotational angiography use is analysed. The case examples of virtual applications use on various anatomic areas are described. Informational value of routine and 3-D angiography is comparatively evaluated. The preferences of modern technology is showed.


Punicalin alleviates TNF-α- and IL-1β-induced chondrocyte dysfunction and cartilage metabolism via mediating FOXO3 signaling axis.
Forkhead box O3 (FOXO3) transcription factor is involved in chondral homeostasis of normal, aging and osteoarthritis (OA) cartilage. At present, we aimed to investigate whether FOXO3 is a target of punicalin to prevent IL-1β- and TNF-α-induced chondrocyte dysfunction in vitro and in vivo models. Cell and mouse models of chondrocyte dysfunction were established to determine the pharmacological value of hydrolyzable tannin, punicalin, which was extracted from the pomegranate. FOXO3 protein levels in the nucleus and cytoplasm were analysed using western blot. Safranine O staining was performed to evaluate the expansion of growth plate and chondrocyte differentiation in IL-1β- and TNF-α-treated mice. In IL-1β- and TNF-α-treated chondrocytes and mice, IL-1β and TNF-α evoked phosphorylation and nucleocytoplasmic shuttling of FOXO3, as well as reduced FOXO3 expression levels in the nucleus. However, punicalin treatment repressed FOXO3 phosphorylation and cytoplasmic transfer. Punicalin treatment improved IL-1β and TNF-α-induced growth inhibition and apoptosis of chondrocyte and the abnormal expansion of growth plate and hypertrophic zone. Moreover, punicalin could maintain the normal phenotype of chondrocyte via mediating multiple gene expression. Punicalin showed a beneficial effect on IL-1β- and TNF-α-stimulated chondrocytes and cartilaginous metabolic disorders via preserving the transcriptional activity of FOXO3. PRACTICAL APPLICATIONS: Our study presents a prospective adjuvant therapeutic drug, punicalin, to prevent inflammation-related cartilage injury and chondrocyte dysfunction.


Low antigenicity of the polysaccharide region of Helicobacter pylori lipopolysaccharides derived from tumors of patients with gastric cancer.
We have examined the antibody response to Helicobacter pylori lipopolysaccharide (LPS) during natural infection in humans. The sera of over 70% of H. pylori-infected individuals were found to contain immunoglobulin G antibodies against the LPS fractions isolated from smooth strains of H. pylori but not against those derived from rough strains, as determined by enzyme-linked immunosorbent assay. These results taken together with the immunoblot data indicated that the polysaccharide region of H. pylori LPS is antigenic in humans. However, the antigenicity of the polysaccharide varied, depending on the strain. We found that smooth H. pylori strains isolated from the tumors of patients with gastric cancer showed significantly lower antigenicity than smooth strains derived from patients with chronic gastritis and gastric and duodenal ulcers. The results suggest that the levels of antigenicity of the polysaccharide region of H. pylori LPS in humans correlate with the nature of the gastroduodenal diseases and that they allow a particular distinction to be made between gastric cancer and other gastroduodenal diseases, especially chronic gastritis.


Furaldehyde substrate specificity and kinetics of Saccharomyces cerevisiae alcohol dehydrogenase 1 variants.
A previously discovered mutant of Saccharomyces cerevisiae alcohol dehydrogenase 1 (Adh1p) was shown to enable a unique NADH-dependent reduction of 5-hydroxymethylfurfural (HMF), a well-known inhibitor of yeast fermentation. In the present study, site-directed mutagenesis of both native and mutated ADH1 genes was performed in order to identify the key amino acids involved in this substrate shift, resulting in Adh1p-variants with different substrate specificities. In vitro activities of the Adh1p-variants using two furaldehydes, HMF and furfural, revealed that HMF reduction ability could be acquired after a single amino acid substitution (Y295C). The highest activity, however, was reached with the double mutation S110P Y295C. Kinetic characterization with both aldehydes and the in vivo primary substrate acetaldehyde also enabled to correlate the alterations in substrate affinity with the different amino acid substitutions. We demonstrated the key role of Y295C mutation in HMF reduction by Adh1p. We generated and kinetically characterized a group of protein variants using two furaldehyde compounds of industrial relevance. Also, we showed that there is a threshold after which higher in vitro HMF reduction activities do not correlate any more with faster in vivo rates of HMF conversion, indicating other cell limitations in the conversion of HMF.


Unexpected medical undergraduate simulation training (UMUST): can unexpected medical simulation scenarios help prepare medical students for the transition to foundation year doctor?
Preparing medical students with the skills necessary to deal with emergency situations as junior doctors can be challenging due to the complexities of creating authentic 'real life' experiences in artificial environments. The following paper is an evaluation of the UMUST (Unexpected Medical Undergraduate Simulation Training) project; a high-fidelity simulation based training programme designed to emulate the experience of dealing with medical emergencies for final year medical students preparing for practice as Foundation Year trainees. Final year medical students from Liverpool University who undertake their clinical placements at Blackpool Teaching Hospitals NHS Foundation Trust and St. Helens & Knowsley Teaching Hospitals NHS Trust were randomly allocated into groups and took part in a series of four unexpected simulation based scenarios. At the beginning of the week in which the scenarios ran, participants were issued with a hospital bleep which they carried with them during their placement. At an unknown time to them, the participants were bleeped to attend a simulated emergency scenario, and on arrival to the Clinical Skills and Simulation facility, members of the education team undertook a standardised simulation scenario. Each session was recorded on video which the participants subsequently watched as part of a debriefing process. An assessment tool was developed to gauge whether the participants made progress in their learning over the course of the four sessions. Focus groups were held with the participants in order to evaluate their experience of the programme, and questionnaires were later distributed to all participants once they had begun working as a Foundation Year trainee. The questionnaires asked them how relevant UMUST was in preparing them for dealing with medical emergencies. The questionnaires and the focus groups clearly showed that the doctors felt like UMUST was very valuable in preparing them to work as junior doctors. They had enjoyed taking part in UMUST and thought was a realistic and useful part of their undergraduate training. The feedback from the focus groups and the subsequent questionnaires clearly demonstrate that participants felt the UMUST programme helped to prepare them as junior doctors in terms of dealing with emergency situations.


Cortisol, but not intranasal insulin, affects the central processing of visual food cues.
Stress glucocorticoids and insulin are important endocrine regulators of energy homeostasis, but little is known about their central interaction on the reward-related processing of food cues. According to a balanced group design, healthy food deprived men received either 40IU intranasal insulin (n=13), 30mg oral cortisol (n=12), both (n=15), or placebo (n=14). Acoustic startle responsiveness was assessed during presentation of food and non-food pictures. Cortisol enhanced startle responsiveness during visual presentation of "high glycemic" food pictures, but not during presentation of neutral and pleasant non-food pictures. Insulin had no effect. Based on the "frustrative nonreward" model these results suggest that the reward value of high glycemic food items is specifically increased by cortisol.


Platelet-Rich Plasma Augments Subcision in Atrophic Acne Scars: A Split-Face Comparative Study.
Acne has a prevalence of 90% among adolescents. Facial scarring affects 75% of patients. Autologous platelet-rich plasma (PRP) is a novel treatment option for acne scar management and can be used as an adjuvant to acne scar revision procedures. Owing to its remodeling properties, PRP when used as an adjuvant reduces the total number of sittings. To compare efficacy of autologous PRP and subcision against subcision alone in acne scars. It was an experimental analytical study conducted over a period of 2 years. Forty patients successfully completed the trial. It was a split-face study in which the right side of the face was the study side where autologous PRP was injected into each scar after performing subcision. The left side of the face was the control side where only subcision was performed. Digital photographs were taken at every sitting. Analysis was performed using the Wilcoxon signed-rank test and Mann-Whitney tests in SPSS software. Platelet-rich plasma and subcision showed greater improvement (32.08%) in postacne scars as compared to subcision alone (8.33%). Rolling acne scars responded greatest (39.27%) followed by box-type scars (33.88%). Platelet-rich plasma and subcision act synergistically to improve the appearance of acne scars.


Outcomes of delivery room resuscitation of bradycardic preterm infants: A retrospective cohort study of randomised trials of high vs low initial oxygen concentration and an individual patient data analysis.
To determine whether hospital mortality (primary outcome) is associated with duration of bradycardia without chest compressions during delivery room (DR) resuscitation in a retrospective cohort study of randomized controlled trials (RCTs) in preterm infants assigned low versus high initial oxygen concentration. Medline and EMBASE were searched from 01/01/1990 to 12/01/2020. RCTs of low vs high initial oxygen concentration which recorded serial heart rate (HR) and oxygen saturation (SpO2) during resuscitation of infants <32 weeks gestational age were eligible. Individual patient level data were requested from the authors. Newborns receiving chest compressions in the DR and those with no recorded HR in the first 2 min after birth were excluded. Prolonged bradycardia (PB) was defined as HR < 100 bpm for ≥2 min. Individual patient data analysis and pooled data analysis were conducted. Data were collected from 720 infants in 8 RCTs. Neonates with PB had higher odds of hospital death before [OR 3.8 (95% CI 1.5, 9.3)] and after [OR 1.7 (1.2, 2.5)] adjusting for potential confounders. Bradycardia occurred in 58% infants, while 38% had PB. Infants with bradycardia were more premature and had lower birth weights. The incidence of bradycardia in infants resuscitated with low (≤30%) and high (≥60%) oxygen was similar. Neonates with both, PB and SpO2 < 80% at 5 min after birth had higher odds of hospital mortality. [OR 18.6 (4.3, 79.7)]. In preterm infants who did not receive chest compressions in the DR, prolonged bradycardia is associated with hospital mortality.


In-depth Characterization of Firefly Luciferase as a Reporter of Circadian Gene Expression in Mammalian Cells.
Firefly luciferase (Fluc) is frequently used to report circadian gene expression rhythms in mammalian cells and tissues. During longitudinal assays it is generally assumed that enzymatic substrates are in saturating excess, such that total bioluminescence is directly proportional to Fluc protein level. To test this assumption, we compared the enzyme kinetics of purified luciferase with its activity in mammalian cells. We found that Fluc activity in solution has a lower Michaelis constant (Km) for luciferin, lower temperature dependence, and lower catalytic half-life than Fluc in cells. In consequence, extracellular luciferin concentration significantly affects the apparent circadian amplitude and phase of the widely used PER2::LUC reporter in cultured fibroblasts, but not in SCN, and we suggest that this arises from differences in plasma membrane luciferin transporter activity. We found that at very high concentrations (>1 mM), luciferin lengthens circadian period, in both fibroblasts and organotypic SCN slices. We conclude that the amplitude and phase of circadian gene expression inferred from bioluminescence recordings should be treated with some caution, and we suggest that optimal luciferin concentration should be determined empirically for each luciferase reporter and cell type.


Direct drug cocktail analyses using microscale vortex-assisted electroporation.
Combination therapy has become one of the leading approaches for treating complex diseases because it coadministers clinically proven drugs to concurrently target multiple signaling pathways of diseased cells. Identification of synergic drug combinations at their respective effective doses without unwanted accumulative side effects is the key to success for such therapy. In this work, we demonstrate the feasibility of the vortex-assisted microfluidic electroporation system for direct drug cocktail analyses where drug substances were individually delivered into cytosols in a sequential and dosage-controlled manner. Through quantitative analyses, the synergic combinational dosage ratios of the chemotherapeutic drug and the anticancer flavonoid were identified. When integrated with high-throughput label-free rare cell purification techniques, the presented system has the potential for development of personalized medicines as the system would be capable of comprehensively assessing drug combinations directly on patients' cellular samples.


Neuronal expression of the glutamate transporter GLT-1 in hippocampal microcultures.
To address the question of the relative contributions of glial and neuronal glutamate transport in the vertebrate CNS, we studied the distribution of forebrain glutamate transporters in rat hippocampal microcultures, a preparation in which physiological functions of glutamate transporters have been well characterized. Two of the three transporters, GLAST (EAAT1) and EAAC1 (EAAT3), are localized to microculture glia and neurons, respectively, as expected. However, we find strong immunoreactivity for the third glutamate transporter GLT-1 (EAAT2), a putatively glial transporter, in microculture neurons and in a small subset of microculture glia. Indistinguishable immunohistochemical staining patterns for GLT-1 were obtained with antibodies directed against both the N terminal and C terminal of the GLT-1 protein. Double-labeling experiments suggest that neuronal GLT-1 protein is primarily localized to the dendrites of excitatory neurons. Neuronal electrogenic transport currents in response to D-aspartate applications were occluded by the selective GLT-1 inhibitor dihydrokainate. In contrast, glia exhibited a larger transporter current density than did neurons, and the glial transport current was less sensitive to dihydrokainate. Neuronal transport currents were potentiated less than were glial currents when the chaotropic anion thiocyanate was substituted for gluconate in the whole-cell recording pipette, consistent with the previously reported lower anion permeability of EAAC1 and GLT-1 compared with that of GLAST. After microculture glia were rendered nonviable, excitatory autaptic currents (EACs) were prolonged in the presence of dihydrokainate, suggesting that neuronal GLT-1 is capable of participating in the clearance of synaptically released glutamate. Our results suggest that the initially proposed characterization of GLT-1 as a purely glial transporter is too simplistic and that under certain conditions functional GLT-1 protein can be expressed in brain neurons. The study suggests that changes in GLT-1 levels that occur with pathology or experimental manipulations cannot be assumed to be glial.


Gemcitabine versus gemcitabine/carboplatin in advanced non-small cell lung cancer: preliminary findings in a phase III trial of the Swedish Lung Cancer Study Group.
Gemcitabine is an active agent in non-small cell lung cancer, with single-agent treatment producing response rates of approximately 20% and median survivals of approximately 7 to 9 months. In a pilot trial in advanced non-small cell lung cancer, the gemcitabine/carboplatin combination produced a response rate of 43% and median survival of 12 months with good tolerability. Preliminary results of a phase III trial comparing gemcitabine alone with gemcitabine/carboplatin in 332 patients with stage IIIB or IV non-small cell lung cancer are now available. Patients were randomized to receive gemcitabine 1,250 mg/m(2) on days 1 and 8 every 21 days or the same gemcitabine regimen plus carboplatin at an area under the concentration-time curve of 5 mg/mL/min on day 1 for a maximum of six cycles. Hematologic toxicity was more common in the combination arm; grade 4 thrombocytopenia occurred in 23.5% v 5.3% of patients, but infrequently resulted in clinical complications. Nonhematologic toxicity was moderate and similar in frequency in the combination and gemcitabine arms (25% and 28%, respectively). Among 275 patients, overall response rates were 30% (2% complete response and 28% partial response) in the combination arm and 12% (all partial responses) in the gemcitabine arm. Median time to disease progression was 6 months in the combination arm and 4 months in the gemcitabine arm. Median survival in the study population was 9 months, a promising finding given the high proportion of elderly patients in the study (37% >/= 70 years of age). Full mature results of the trial, including comparative survival results and data on quality of life, are awaited.


Multiple endocrine neoplasia type 1: a 10-year prospective screening study in four kindreds.
A total of 80 individuals in 4 kindreds with multiple endocrine neoplasia type 1 (MEN 1) have been subjected to repeated biochemical screening during a 10-yr period with the principal aim being to analyze characteristics of the developing pancreatic lesion. Age at presentation of the MEN 1 trait averaged 18 yr in 7 previously unaffected individuals, and this effect of the screening procedure represented a lowering by almost 2 decades. Pancreatic endocrine involvement was recognized at a mean age of 25 yr and constituted the presenting lesion in a majority of the patients. A standardized meal test and basal values of serum pancreatic polypeptide, insulin, proinsulin, and gastrin were the most efficient markers for the pancreatic lesion and preceded signs of pancreatic tumors upon radiological examinations by a mean of 3.5 yr. A 75% penetrance of the islet cell disease and 90% for primary hyperparathyroidism within the affected individuals equalled the prevalences reported in autopsy studies. Two of the kindreds showed signs of intrafamilial homogeneity with respect to the profile of peptide excess (P less than 0.05) and considerable discrepancy in the malignant potential of the pancreatic lesions. The results of early detection and surgical intervention of the pancreatic tumors in MEN 1 suggested an impact on morbidity, while any effect on the mortality of these individuals remains to be clarified.


Prevalence and socioeconomic impact of upper gastrointestinal disorders in the United States: results of the US Upper Gastrointestinal Study.
This study examined the prevalence of upper gastrointestinal (GI) symptoms and symptom groupings and determined impact on disability days in a nationally representative US sample. A telephone survey of 21,128 adults was conducted including questions about the presence of upper GI symptoms during the past 3 months. Respondents were categorized as symptomatic (ie, reported GI symptoms once per month) or asymptomatic. The survey included questions about missed work, leisure activity, or household activity days. Symptom groupings were identified by using factor analysis, and cluster analysis was used to assign respondents into distinct groups on the basis of these symptom groupings. The prevalence of an average of 1 or more upper GI symptoms during the past 3 months was 44.9%. The most common symptoms experienced during the past 3 months were early satiety, heartburn, and postprandial fullness. Factor analysis identified 4 symptom groupings: (1) heartburn/regurgitation; (2) nausea/vomiting; (3) bloating/abdominal pain; and (4) early satiety/loss of appetite. Five respondent clusters were identified; the largest clusters were primarily early satiety/fullness (44%) and gastroesophageal reflux disease-like symptoms (28%). Two small clusters reflected nausea and vomiting (7%) and a heterogeneous symptom profile (4%). Symptomatic respondents reported significantly more missed work, leisure, and household activity days than asymptomatic respondents (all P < .0001). Factor analysis separated GI symptoms into groupings reflecting gastroesophageal reflux disease and dyspepsia: early satiety, postprandial fullness, and loss of appetite; bloating and abdominal pain/discomfort; and nausea and vomiting. These upper GI symptoms were associated with significant loss of work and activity days.


Online radiographic survey of midshaft clavicular fractures: no consensus on treatment for displaced fractures.
The choice of treatment for midshaft clavicular fractures is not straightforward, but depends on fracture characteristics such as comminution, angulation and displacement. An online survey was conducted amongst trauma and orthopaedic surgeons to determine the preferred treatment for midshaft clavicular fractures, based on anteroposterior radiographs, for 17 randomly selected displaced or comminuted midshaft clavicular fractures. The background and experience of the respondents were documented. Data were analyzed using a Generalized Estimating Equations (GEE) model. The 102 respondents preferred non-operative treatment more frequently for displaced fractures than for comminuted fractures (OR 3.24, 95% CI 2.55-4.12). Locking plate fixation was more often preferred over other surgical modalities for comminuted than for displaced fractures (OR 1.50, 95% CI 1.17-1.91). In clinical practice, there is no consensus between surgeons on the choice of treatment for displaced or comminuted midshaft clavicular fractures. This lack of agreement calls for evidence-based treatment guidelines for these fractures.


"Done more for me in a fortnight than anybody done in all me life." How welfare rights advice can help people with cancer.
In the UK many people with cancer and their carers do not have easy access to the welfare benefits to which they are entitled adding further strain to the process of dealing with cancer. It is estimated that nine out of ten cancer patients' households experience loss of income as a direct result of cancer, which, due to its socio-economic patterning disproportionately affects those most likely to be financially disadvantaged. In the UK proactive welfare rights advice services accessed via health care settings significantly increase benefit entitlement among people with health problems and this paper reports on a qualitative study examining the impact of a welfare rights advice service specifically designed for people affected by cancer and their carers in County Durham, North East England (UK). Twenty two men and women with cancer or caring for someone with cancer who were recipients of welfare rights advice aged between 35 and 83 were recruited from a variety of health care and community settings. Semi-structured interviews were undertaken and analysed using the Framework method. Most of the participants experienced financial strain following their cancer diagnosis. Participants accessed the welfare rights service in a variety of ways, but mainly through referral by other professionals. The additional income generated by successful benefit claims was used in a number of ways and included offsetting additional costs associated with cancer and lessening the impact of loss of earnings. Overall, receiving welfare rights advice eased feelings of stress over financial issues at a time when participants were concerned about dealing with the impact of cancer. Lack of knowledge about benefit entitlements was the main barrier to accessing benefits, and this outweighed attitudinal factors such as stigma and concerns about benefit fraud. Financial strain resulting from a cancer diagnosis is compounded in the UK by lack of easy access to information about benefit entitlements and assistance to claim. Proactive welfare rights advice services, working closely with health and social care professionals can assist with the practical demands that arise from dealing with the illness and should be considered an important part of a holistic approach to cancer treatment.


Behavioural interventions promoting condom use among female sex workers in sub-Saharan Africa: a systematic review.
The objective of this review was to provide an overview of behavioural interventions promoting condom use amongst female sex workers (FSW) in sub-Saharan Africa. A search of four electronic bibliographic databases from 1990 to September 2016 was carried out. The search was limited to articles published in English. Studies which evaluated behavioural interventions to increase condom use among FSWs were selected and reviewed. Data were extracted on effectiveness, condom use, intervention content, and process outcomes. A total of 20 eligible articles describing 18 interventions in sub-Saharan Africa on HIV prevention with condom use as an outcome measure were identified. Most of the behavioural interventions incorporated a combination of approaches: health education by peers, health workers and project staff, and activities by brothel owners and brothel managers. Most studies showed effectiveness of these interventions on condom use with paying clients. Five studies measuring condom use with regular non-paying partners recorded less consistent condom use with these partners. This review illustrates the existence of sufficient evidence showing the effectiveness of behavioural interventions targeting correct and consistent condom use by FSWs.


Worldwide and Country-Specific Impact of the COVID-19 Pandemic on Heart Transplantation Volumes: A Longitudinal Analysis of 2020 and 2021.
COVID-19 pandemic hampered operational efficiency of heart transplant (HT) programs worldwide. Little is known about the global and country-specific changes in HT volumes during the pandemic years 2020-2021. We aimed to describe the global and country-level impact of the COVID-19 pandemic on HT volumes in 2020-2021. This is a cross-sectional study of the Global Observatory on Donation and Transplantation, including the years 2019-2021. Among 60 countries that reported HT data in the years 2019-2020, we analyzed 52 countries with ≥1 transplant during each year. Overall, the number of HTs decreased during 2020 by 9.3% (1.82 to 1.65 PMP). While 75% (n = 39/52) of countries experienced a decrease in HT volumes in 2020, volumes were maintained/increased in the remaining countries. Countries with maintained HT volumes had a higher organ donation rate in 2020 compared to those with decreased volumes (P = 0.03), the only significant predictor of change in HT volumes (P = 0.005). In 2021, a 6.6% recovery from the previous year's drop in global HT rate was noticed, reaching 1.76 HT PMP. Only 1 in 5 countries with reduced volumes in 2020 recovered their baseline volumes in 2021. Only 30.8% of countries with maintained volumes in 2020 had continued growth in HT volumes in 2021. The latter group encompassed the United States of America, the Netherlands, Poland, and Portugal. Further work should define underlying causes of this heterogeneity in HT volume during the pandemic. Identifying policies and practices that helped certain countries mitigate the effect of the pandemic on HT activities may help other countries during similar health crises in the future.


Effect of PTH and 1,25(OH)2D3 on renal 25(OH)D3 metabolism, adenylate cyclase, and protein kinase.
The purpose of these studies was to characterize the action of PTH and 1,25(OH)2D3 on the renal metabolism of 25(OH)D3 to 1,25(OH)2D3 and 24,25(OH)2D3. Renal metabolism of 25(OH)D3, adenylate cyclase, and protein kinase activity were measured using isolated renal slices from rats fed a vitamin D-deficient, low-calcium diet and thyroparathyroidectomized. PTH added to renal slices for 4 h in vitro maximally increased 1,25(OH)2D3 production by 67% and decreased 24,25(OH)2D3 production by 24% over the concentration range 0.05-5.0 U/ml. Parathyroid hormone (PTH) (0.05 U/ml) added to renal slices for 5 min produced a significant increase in tissue cAMP and a near-maximal increase in cAMP-dependent protein kinase activity. Preincubation of renal slices with 50 nM 1,25(OH)2D3 decreased renal 1,25(OH)2D3 production by 26% and increased 24,25(OH)2D3 production by 55%. 1,25(OH)2D3 also blocked the effect of PTH (5.0 U/ml) on renal 25(OH)D3 metabolism. However, PTH-stimulated adenylate cyclase and protein kinase activity was not blocked by preincubation with 1,25(OH)2D3. These studies demonstrate that PTH may act directly on the kidney to modulate renal 25(OH)D3 metabolism and that this action can be inhibited by 1,25(OH)2D3. This inhibition by 1,25(OH)2D3 occurs at a site distal to or separate from PTH-stimulated protein kinase activity.


Approaches to modelling irradiation-induced processes in transmission electron microscopy.
The recent progress in high-resolution transmission electron microscopy (HRTEM) has given rise to the possibility of in situ observations of nanostructure transformations and chemical reactions induced by electron irradiation. In this article we briefly summarise experimental observations and discuss in detail atomistic modelling of irradiation-induced processes in HRTEM, as well as mechanisms of such processes recognised due to modelling. Accurate molecular dynamics (MD) techniques based on first principles or tight-binding models are employed in the analysis of single irradiation-induced events, and classical MD simulations are combined with a kinetic Monte Carlo algorithm to simulate continuous irradiation of nanomaterials. It has been shown that sulphur-terminated graphene nanoribbons are formed inside carbon nanotubes as a result of an irradiation-selective chemical reaction. The process of fullerene formation in HRTEM during continuous electron irradiation of a small graphene flake has been simulated, and mechanisms driving this transformation analysed.


What non-alcoholic fatty liver disease has got to do with obstructive sleep apnoea syndrome and viceversa?
Non-alcoholic fatty liver disease (NAFLD) and obstructive sleep apnoea syndrome (OSAS) are common conditions, frequently encountered in patients with obesity and/or metabolic syndrome. NAFLD and OSAS are complex diseases that involve an interaction of several intertwined factors. Several lines of evidence lend credence to an immune system derangement in these patients, i.e. the low grade chronic inflammation status, reckoned to be the most important factor in causing and maintaining these two illnesses. Furthermore, it is emphasized the main role of spleen involvement, as a novel mechanism. In this review the contribution of the visceral adiposity in both NAFLD and OSAS is stressed as well as the role of intermittent hypoxia. Finally, a post on the prevention of systemic inflammation is made.


Musculoskeletal disorders of the neck and shoulders in female sewing machine operators: prevalence, incidence, and prognosis.
To assess the occurrence and persistence of two restrictively defined neck-shoulder disorders among sewing machine operators. To assess factors associated with the development of neck-shoulder disorder and prognostic factors for remaining a case, when disorders were already present. In an initial group of 243 sewing machine operators, 178 were followed up for 2 years. At baseline and at 1 and 2 years follow up the participants underwent a clinical examination of the neck and arms and filled in a questionnaire about current musculoskeletal complaints. Clinical criteria for two main neck-shoulder disorders were defined: rotator cuff tendinitis and myofascial pain syndrome. A baseline control group consisted of 357 women with varied non-repetitive work. At baseline the overall prevalence of myofascial pain syndrome and rotator cuff tendinitis was 15.2% and 5.8% among sewing machine operators compared with 9.0% and 2.2%, respectively, among controls. The presence of the disorders was strongly associated with a self perception of poor general health. Although myofascial pain syndrome showed a U shaped association with years as a sewing machine operator, rotator cuff tendinitis was absent among the newest recruits and present among 15% of the women with more than 20 years as a sewing machine operator. Besides years as a sewing machine operator, the risk of having a neck-shoulder disorder at baseline was significantly associated with high stress (prevalence ratio (PR)=2.54; 95% confidence interval (95% CI) 1.28 to 5.05) when adjusted for age, body mass index, smoking, living alone with children, job strain, and social support from colleagues and supervisors. Only one of 13 participants with rotator cuff tendinitis at baseline recovered during follow up. Myofascial pain syndrome showed a much more fluctuating tendency. Low social support (RR 3.72; 95% CI 1.22 to 11.30) and smoking (RR 3.93; 95% CI 1.33 to 11.58) were associated with the development of neck-shoulder disorders, which was also associated with neck-shoulder pain score and living alone with children. Rotator cuff tendinitis showed a higher degree of persistence than myofascial pain syndrome. Both disorders highly influenced the perception of general health. Women who lived alone with children, were smokers, or experienced low support from colleagues and supervisors had a higher risk of contracting a neck-shoulder disorder.


Induction of remission in female rheumatoid arthritis patients is associated with stabilization of myocardial abnormalities: a prospective cardiac magnetic resonance follow-up study.
Objectives: To study whether female patients with active rheumatoid arthritis (RA) have myocardial abnormalities and whether progression of myocardial involvement can be attenuated by disease-modifying anti-rheumatic drugs (DMARDs).Method: Cardiac magnetic resonance (cMR; 1.5 or 3.0 T), including late gadolinium enhancement (LGE), T1 relaxation time, and ventricular functions, was performed in 30 patients with untreated active early RA starting first DMARDs, and 28 patients with chronic RA with inadequate response to conventional synthetic DMARDs starting biological DMARDs. cMR was repeated in RA patients 1 year later. cMR was conducted once in 22 fibromyalgia (FM) subjects and in 35 healthy volunteers serving as controls. All subjects were non-smoking females without coronary heart disease, heart failure, or diabetes.Results: Compared with controls, 58 RA patients had slightly lower ventricular function, although in the normal range, and longer T1 time at baseline. None of the FM subjects had LGE, but it was frequent in RA (67%). During the 1 year DMARD treatment, Disease Activity Score based on 28-joint count-C-reactive protein declined, ventricular functions tended to improve, but the number of patients with LGE remained unchanged. However, the number of LGE-positive heart segments either decreased or stayed the same in 91% of RA patients. In early RA patients, achieving tight remission was associated with LGE stabilization, after adjustment for age, metabolic syndrome, baseline inflammatory activity, and leisure-time physical activity.Conclusion: Treatment targeted to tight remission in early stages of RA seems to be important to prevent not only joint damage but also myocardial abnormalities.


[The forensic medical expertise in trauma of male genitals].
A damage inflicted to genitals organs (GO) affects the copulative functions, the sperm fertilizing ability and, as a whole, the reproductive function. Injuries to the external genital organs (EGO) due to immediate trauma are more often the case in the forensic medical practice; they are less often encountered in operative interventions in organs, which are close to the prostate or those having immediate direct links with the testicles or with the cavernous bodies of the penis, i.e. in case of an indirect trauma or an mediated effect on the functions of GO. Even if there is a smallest damage to the EGO region, the man needs to consult the urologist in the out-patient or in-patient medical institution. It is always necessary to asses a degree of severity to the health both by signs of duration to health disorders and by an outcome of injury with regard for trauma severity and possible impact on the reproductive function.


Glycosylation motifs that direct arabinogalactan addition to arabinogalactan-proteins.
Hydroxyproline (Hyp)-rich glycoproteins (HRGPs) participate in all aspects of plant growth and development. HRGPs are generally highly O-glycosylated through the Hyp residues, which means carbohydrates help define the interactive molecular surface and, hence, HRGP function. The Hyp contiguity hypothesis predicts that contiguous Hyp residues are sites of HRGP arabinosylation, whereas clustered noncontiguous Hyp residues are sites of galactosylation, giving rise to the arabinogalactan heteropolysaccharides that characterize the arabinogalactan-proteins. Early tests of the hypothesis using synthetic genes encoding only clustered noncontiguous Hyp in the sequence (serine [Ser]-Hyp-Ser-Hyp)(n) or contiguous Hyp in the series (Ser-Hyp-Hyp)(n) and (Ser-Hyp-Hyp-Hyp-Hyp)(n) confirmed that arabinogalactan polysaccharide was added only to noncontiguous Hyp, whereas arabinosylation occurred on contiguous Hyp. Here, we extended our tests of the codes that direct arabinogalactan polysaccharide addition to Hyp by building genes encoding the repetitive sequences (alanine [Ala]-proline [Pro]-Ala-Pro)(n), (threonine [Thr]-Pro-Thr-Pro)(n), and (valine [Val]-Pro-Val-Pro)(n), and expressing them in tobacco (Nicotiana tabacum) Bright-Yellow 2 cells as fusion proteins with green fluorescent protein. All of the Pro residues in the (Ala-Pro-Ala-Pro)(n) fusion protein were hydroxylated and consistent with the hypothesis that every Hyp residue was glycosylated with arabinogalactan polysaccharide. In contrast, 20% to 30% of Pro residues remained non-hydroxylated in the (Thr-Pro-Thr-Pro)(n), and (Val-Pro-Val-Pro)(n) fusion proteins. Furthermore, although 50% to 60% of the Hyp residues were glycosylated with arabinogalactan polysaccharide, some remained non-glycosylated or were arabinosylated. These results suggest that the amino acid side chains of flanking residues influence the extent of Pro hydroxylation and Hyp glycosylation and may explain why isolated noncontiguous Hyp in extensins do not acquire an arabinogalactan polysaccharide but are arabinosylated or remain non-glycosylated.


Ketamine spares morphine consumption after transthoracic lung and heart surgery without adverse hemodynamic effects.
Thoracotomy is associated with severe pain. Large doses of morphine can depress respiratory drive and compromise hemodynamic stability. Ketamine reduces hyperalgesia, prevents opioid tolerance and resistance and lowers morphine consumption. At sub-anesthetic (< or = 500 microg/kg) doses, ketamine rarely produces undesirable hemodynamic alterations. We hypothesized that by combining a sub-anesthetic dose of ketamine with morphine, we could effectively control pain with less morphine and minimize drowsiness, while maintaining safe hemodynamic and respiratory parameters. Sequential patients undergoing anterolateral thoracotomy for MIDCAB, lung tumor resection, or median sternotomy for OPCAB were randomized to one of the two intravenous patient-controlled analgesia (IV-PCA) protocols. MO-only patients received 1.5 mg MO bolus, and MK patients received 1.0 mg MO+5 mg ketamine/bolus, both with a 7 min lockout time. IV-PCA was initiated when the patient was sufficiently awake (> or = 5/10 VAS) and rated pain > or = 5/10 on a 0-10 VAS. Rescue intramuscular diclofenac 75 mg was available. Follow-up of respiratory, hemodynamic and pain statuses lasted 72 h. Fifty-eight patients completed the 6-month study. Heart rate and blood pressures were identically stable in both groups. Respiratory rate and pulse oximetry were higher (P < 0.05) in the MK than in the MO group. MO patients (n = 28) used twice (2.0 +/-2.3 mg/patient/h) the amount of morphine compared to MK patients (n = 30, 1.0 +/- 1.4 mg/patient/h, P < 0.05). Thirty-six hours after starting PCA, 10 MO patients still required IV-PCA compared to 5 MK patients (P < 0.05). Diclofenac was used 70% more in MO than in MK patients. MO patients suffered more postoperative nausea and vomiting. No patients had hallucinations. The concomitant use of sub-anesthetic ketamine plus two-thirds the standard MO dose following thoracotomy, MIDCAB or OPCAB resulted in lower pain scores, reduced MO consumption and shorter postoperative IV-PCA dependence. These advantages were associated with cardiovascular stability and even better respiratory parameters.


Dynamics of three organic acids (malic, acetic and succinic acid) in sunflower exposed to cadmium and lead.
Sunflower (Helianthus annuus L.) has been considered as a good candidate for bioaccumulation of heavy metals. In the present study, sunflower was used to enrich the cadmium and lead in sand culture during 90 days. Biomass, Cd and Pb uptake, three organic acids and pH in cultures were investigated. Results showed that the existence of Cd and Pb showed different interactions on the organic acids exudation. In single Cd treatments, malic and acetic acids in Cd10 showed an incremental tendency with time. In the mixed treatments of Cd and Pb, malic acids increased when 10 and 40 mg x L(-1) Cd were added into Pb50, but acetic acids in Pb50 were inhibited by Cd addition. The Cd10 supplied in Pb10 stimulated the secretion of malic and succinic acids. Moreover, the Cd or Pb uptake in sunflower showed various correlations with pH and some organic acids, which might be due to the fact that the Cd and Pb interfere with the organic acids secretion in rhizosphere of sunflower, and the changes of organic acids altered the form and bioavailability of Cd and Pb in cultures conversely.


[Essential public health functions in the nursing curriculum at Federal University of Rio Grande do Sul].
The Pan American Health Organization referential of Essential Public Health Functions (EPHF) was developed to improve public health performance, which depends on professional education. A case study with the objective of identifying the insertion of EPHF in the curriculum of the nursing undergraduate course at Federal University of Rio Grande do Sul was developed. Keywords were defined from the definitions of the eleven EPHF. These were searched for in the class plans of the curriculum courses. Ten EPHF were identified, with major occurrence of the human health resources development and training in public health. Contemplating more intensively some of the EPHF, the curriculum under study should propitiate the education of nurses capable of contributing towards their performance. It is necessary to insert the other EPHF into the curriculum, contributing to the excellent performance of public health by the professionals educated in this institution.


SGLT-2 Inhibitors and Cardiovascular Risk in Diabetes Mellitus: A Comprehensive and Critical Review of the Literature.
Diabetes mellitus is a major cardiovascular risk factor. Despite the vast pharmaceutical armamentarium, current therapeutic options for the treatment of type 2 diabetes mellitus were unable to provide consistent cardiovascular benefits, apart for metformin. The newest antidiabetic class of drugs, the SGLT-2 inhibitors, seem to provide significant survival benefits. The aim of this review is to present available data that will clarify whether SGLT-2 inhibitors could precipitate in reducing cardiovascular risk in diabetes mellitus. A comprehensive literature search was performed to identify available data from clinical and experimental studies evaluating the impact of SGLT-2 inhibitors in cardiovascular risk factors and outcomes. Along with a mild antihyperglycemic effect, SGLT-2 inhibitors seem to possess multi-dimensional properties, affecting positively several recognized cardiovascular risk factors such as increased blood pressure, arterial stiffness, body weight and dyslipidemia. Furthermore, preliminary data point towards additional benefits, including reduction of albuminuria in diabetic nephropathy, and reduction of oxidative markers and fatty liver disease in experimental models. The efficacy and safety profile of empagliflozin, an SGLT-2 inhibitor, was evaluated in the EMPA-REG outcome study, and the results were quite impressive. Further credence comes from the analysis of pooled data of SGLT-2 inhibitors trials that have shown similar results. SGLT-2 Inhibitors seem to be related with ameliorating effects on multiple cardiovascular risk factors. The first data from the EMPA-REG outcome study are indeed very promising. Several ongoing cardiovascular studies with this novel class of drugs will shed light and enforce or question current enthusiasm.


Domain-level Identification of Single Prokaryotic Cells by Optical Photothermal Infrared Spectroscopy.
Infrared spectroscopy is used for the chemical characterization of prokaryotes. However, its application has been limited to cell aggregates and lipid extracts because of the relatively low spatial resolution of diffraction. We herein report optical photothermal infrared (O-PTIR) spectroscopy of prokaryotes for a domain-level diagnosis at the single-cell level. The technique provided infrared spectra of individual bacterial as well as archaeal cells, and the resulting aliphatic CH3/CH2 intensity ratios showed domain-specific signatures, which may reflect distinctive cellular lipid compositions; however, there was interference by other cellular components. These results suggest the potential of O-PTIR for a domain-level diagnosis of single prokaryotic cells in natural environments.


[Diflucortolone valerianate (Nerisona) in the treatment of corticoid-sensitive dermatoses in childhood].
The efficacy and tolerance of 0.1% diflucortolone valerate (Nerisona) in cream, ointment and fatty ointment formulations were investigated in a study in which no control preparations were used. The study was conducted in 657 children over a period of three weeks. The results were compared with those of a trial with the same design conducted with 4,878 adults. Our observations were mainly concentrated on the two groups eczema/dermatitis and neurodermatitis (atopic dermatitis). Nearly 90% of the children taking part in the study exhibited these clinical pictures. By far the best therapeutic results were obtained in neurodermatitis. There was only one therapy failure among 198 patients (99.5%). Overall the results obtained in children were markedly better than those recorded for adults.


[Cardiovascular events linked to the new diagnostic categories of diabetes mellitus].
To study the cardiovascular events linked to the new diagnostic categories of diabetes mellitus proposed by the American Diabetes Association. Descriptive cross-sectional study.Setting. Urban health district.Participants. 2248 patients > 14, selected by simple random sampling from the medical records archive. Through the medical records review, patients were classified into those with normal glucaemia, those intolerant to glucose when fasting (IGF) and those with type-2 diabetes mellitus (DM2). Patients were studied for cardiovascular events, ischaemic cardiopathy, cerebrovascular disease and peripheral arteriopathy. The likelihood of such events in patients with IGF and DM2 was compared with normal-glucose patients through logistical regression, adjustment for age, gender, hypertension, hypercholesterolaemia, hypertriglyceridaemia and tobacco dependency. 1677 patients were classified: 1237 with normal glucaemia, 175 with IGF and 265 with DM2. The DM2 patients were more likely to have at least one cardiovascular event (OR, 2.7; 95% CI, 1.7-4.4), ischaemic cardiopathy (OR, 2.0; 95% CI, 1.1-3.6), cerebrovascular disease (OR, 3.0; 95% CI, 1.4-6.6) or peripheral arteriopathy (OR, 2.9; 95% CI, 1.2-7.7). Results reached no statistical significance in patients with IGF, though these were more likely to suffer a cardiovascular event (OR, 1.1; 95% CI, 0.6-2.1), ischaemic cardiopathy (OR, 1.1; 95% CI, 0.3-4.4), cerebrovascular disease (OR, 1.5; 95% CI, 0.5-4.1) or peripheral arteriopathy (OR, 1.2; 95% CI, 0.3-4.6). Patients with DM2 are more likely to present cardiovascular events, a phenomenon not seen in the case of IGF.


Postoperative dressing and management strategies for transtibial amputations: a critical review.
Postamputation management is an important determinant of recovery from amputation. However, consensus on the most effective postoperative management strategies for individuals undergoing transtibial amputation (TTA) is lacking. Dressings can include simple soft gauze dressings, thigh-high rigid cast dressings, shorter removable rigid dressings, and prefabricated pneumatic dressings. Postoperative prosthetic attachments can be added to all but simple soft dressings. These dressings address the need to cleanly cover a fresh surgical wound, but not all postoperative dressings are designed to facilitate the strategic goals of preventing knee contractures, reducing edema, protecting from external trauma, or facilitating early weight bearing. The type of dressing and management strategy often overlap and are certainly interrelated. Current protocols and decisions are based on local practice, skill, and intuition. The current available literature is challenging, and difficulties include variations in healing potential, in comorbidity, in surgical-level selection, in techniques and skill, in experience with postoperative strategies, and with poorly defined outcome criteria. This paper reviews the published literature and compares measures of safety, efficacy, and clinical outcomes of the various techniques. Analysis of 10 controlled studies supported only 4 of the 14 claims cited in uncontrolled, descriptive studies.


Toward a Reliable Collection of Eye-Tracking Data for Image Quality Research: Challenges, Solutions, and Applications.
Image quality assessment potentially benefits from the addition of visual attention. However, incorporating aspects of visual attention in image quality models by means of a perceptually optimized strategy is largely unexplored. Fundamental challenges, such as how visual attention is affected by the concurrence of visual signals and their distortions; whether visual attention affected by distortion or that driven by the original scene only should be included in an image quality model; and how to select visual attention models for the image quality application context, remain. To shed light on the above unsolved issues, designing and performing eye-tracking experiments are essential. Collecting eye-tracking data for the purpose of image quality study is so far confronted with a bias due to the involvement of stimulus repetition. In this paper, we propose a new experimental methodology to eliminate such inherent bias. This allows obtaining reliable eye-tracking data with a large degree of stimulus variability. In fact, we first conducted 5760 eye movement trials that included 160 human observers freely viewing 288 images of varying quality. We then made use of the resulting eye-tracking data to provide insights into the optimal use of visual attention in image quality research. The new eye-tracking data are made publicly available to the research community.


Oncogenic Receptor Tyrosine Kinases Directly Phosphorylate Focal Adhesion Kinase (FAK) as a Resistance Mechanism to FAK-Kinase Inhibitors.
Focal adhesion kinase (FAK) is a major drug target in cancer and current inhibitors targeted to the ATP-binding pocket of the kinase domain have entered clinical trials. However, preliminary results have shown limited single-agent efficacy in patients. Despite these unfavorable data, the molecular mechanisms that drive intrinsic and acquired resistance to FAK-kinase inhibitors are largely unknown. We have demonstrated that receptor tyrosine kinases (RTK) can directly bypass FAK-kinase inhibition in cancer cells through phosphorylation of FAK's critical tyrosine 397 (Y397). We also showed that HER2 forms a direct protein-protein interaction with the FAK-FERM-F1 lobe, promoting direct phosphorylation of Y397. In addition, FAK-kinase inhibition induced two forms of compensatory RTK reprogramming: (i) the rapid phosphorylation and activation of RTK signaling pathways in RTKHigh cells and (ii) the long-term acquisition of RTKs novel to the parental cell line in RTKLow cells. Finally, HER2 +: cancer cells displayed resistance to FAK-kinase inhibition in 3D growth assays using a HER2 isogenic system and HER2+ cancer cell lines. Our data indicate a novel drug resistance mechanism to FAK-kinase inhibitors whereby HER2 and other RTKs can rescue and maintain FAK activation (pY397) even in the presence of FAK-kinase inhibition. These data may have important ramifications for existing clinical trials of FAK inhibitors and suggest that individual tumor stratification by RTK expression would be important to predict patient response to FAK-kinase inhibitors. Mol Cancer Ther; 15(12); 3028-39. ©2016 AACR.


Experiences and effects of psychiatric stigma: Monologues of the stigmatizers and the stigmatized in an African setting.
People with mental illness (PWMI) are faced with a number of social and health-related challenges especially stigma and discrimination which tend to have negative effects on their lives. This paper presents narrative monologues portraying the experiences and effects of psychiatric stigma and discrimination on PWMI in South Africa. These narratives voice out the concerns of the stigmatizers (specifically family members and significant others of PWMI) and the stigmatized in a poetic fashion. The society is still not very sympathetic to the plights of PWMI and this affects their general health and well-being. Traditional beliefs and prejudice still drive public attitude towards PWMI especially in African settings. These narratives presented in a poetic fashion in this paper highlight some salient issues relating to the experience and effects of stigma and the desires of PWMI to be treated with love and respect and helped to lead healthy normal lives.


[Gaze behavior in infancy as an indicator of information processing].
Many studies on perceptual development in infancy use visual attention behaviour as an indicator of intellectual capacities. Decrement of attention with repeated exposure to a visual stimulus (habituation) and recovery of attention to a novel stimulus (dishabituation) are interpreted as reflecting the infant's processing of stimulus information. This information processing interpretation as well as the main empirical habituation-dishabituation procedures are explained. Furthermore, sensory adaptation arguments for habituation and dishabituation are discussed and refuted. Following this critical discussion, empirical evidence for the information processing point of view is given. In this context, investigations are mentioned which indicate substantial correlations between early habituation and dishabituation and later intelligence. These studies support a continuity perspective of mental development from infancy.


Role of β-arrestin-2 in short- and long-term opioid tolerance in the dorsal root ganglia.
G-protein-biased agonists with reduced β-arrestin-2 activation are being investigated as safer alternatives to clinically-used opioids. β-arrestin-2 has been implicated in the mechanism of opioid-induced antinociceptive tolerance. Opioid-induced analgesic tolerance is classically considered as centrally-mediated, but recent reports implicate nociceptive dorsal root ganglia neurons as critical mediators in this process. Here, we investigated the role of β-arrestin-2 in the mechanism of opioid tolerance in dorsal root ganglia nociceptive neurons using β-arrestin-2 knockout mice and the G-protein-biased μ-opioid receptor agonist, TRV130. Whole-cell current-clamp electrophysiology experiments revealed that 15-18-h overnight exposure to 10 μM morphine in vitro induced acute tolerance in β-arrestin-2 wild-type but not knockout neurons. Furthermore, in wild-type neurons circumventing β-arrestin-2 activation by overnight treatment with 200 nM TRV130 attenuated tolerance. Similarly, acute morphine tolerance in vivo in β-arrestin-2 knockout mice was prevented in the warm-water tail-withdrawal assay. Treatment with 30 mg/kg TRV130 s.c. also inhibited acute antinociceptive tolerance in vivo in wild-type mice. Alternately, in β-arrestin-2 knockout neurons tolerance induced by 7-day in vivo exposure to 50 mg morphine pellet was conserved. Likewise, β-arrestin-2 deletion did not mitigate in vivo antinociceptive tolerance induced by 7-day exposure to 25 mg or 50 mg morphine pellet in both female or male mice, respectively. Consequently, these results indicated that β-arrestin-2 mediates acute but not chronic opioid tolerance in dorsal root ganglia neurons and to antinociception in vivo. This suggests that opioid-induced antinociceptive tolerance may develop even in the absence of β-arrestin-2 activation, and thus significantly affect the clinical utility of biased agonists.


Determination of antimycin-A in water by liquid chromatographic/mass spectrometry: single-laboratory validation.
An LC/MS method was developed and validated for the quantitative determination and confirmation of antimycin-A (ANT-A) in water from lakes or streams. Three different water sample volumes (25, 50, and 250 mL) were evaluated. ANT-A was stabilized in the field by immediately extracting it from water into anhydrous acetone using SPE. The stabilized concentrated samples were then transported to a laboratory and analyzed by LC/MS using negative electrospray ionization. The method was determined to have adequate accuracy (78 to 113% recovery), precision (0.77 to 7.5% RSD with samples > or = 500 ng/L and 4.8 to 17% RSD with samples < or = 100 ng/L), linearity, and robustness over an LOQ range from 8 to 51 600 ng/L.


Identification of dynamical hinge points of the L1 ligase molecular switch.
The L1 ligase is an in vitro selected ribozyme that uses a noncanonically base-paired ligation site to catalyze regioselectively and regiospecifically the 5' to 3' phosphodiester bond ligation, a reaction relevant to origin of life hypotheses that invoke an RNA world scenario. The L1 ligase crystal structure revealed two different conformational states that were proposed to represent the active and inactive forms. It remains an open question as to what degree these two conformers persist as stable conformational intermediates in solution, and along what pathway are they able to interconvert. To explore these questions, we have performed a series of molecular dynamics simulations in explicit solvent of the inactive-active conformational switch in L1 ligase. Four simulations were performed departing from both conformers in both the reactant and product states, in addition to a simulation where local unfolding in the active state was induced. From these simulations, along with crystallographic data, a set of four virtual torsion angles that span two evolutionarily conserved and restricted regions were identified as dynamical hinge points in the conformational switch transition. The ligation site visits three distinct states characterized by hydrogen bond patterns that are correlated with the formation of specific contacts that may promote catalysis. The insights gained from these simulations contribute to a more detailed understanding of the coupled catalytic/conformational switch mechanism of L1 ligase that may facilitate the design and engineering of new catalytic riboswitches.


Mortality after pediatric lung transplantation: autopsies vs. clinical impression.
Appreciable mortality accompanies pediatric lung and heart-lung transplantation. The objective of this investigation was to compare the clinical impression of causes of death with autopsy findings in all pediatric lung or heart lung transplant recipients who had an autopsy performed between 1985-2002 at the Children's Hospital of Pittsburgh. Medical records and autopsy findings were reviewed. Thirty recipients with autopsies had 33 transplant procedures: heart-lung (16), double lung (14), repeat lung (2), and repeat heart-lung (1). Perioperative deaths occurred in 8 children, most often precipitated by graft dysfunction. Early deaths (2 weeks-1 year) occurred in 12 children resulting from infection. Late deaths (greater than 1 year) occurred in 10 children. Bronchiolitis obliterans complicated by infection was the major cause of death in these recipients. An autopsy confirmed the clinical impression of cause of death in 29/30 and added significant supplemental information in 16 cases. Unsuspected factors contributing to death included donor lung abnormalities, concurrent infection, and cardiovascular disease. Postmortem examination remains a critical component to augment the understanding of causes of death following pediatric thoracic transplantation.


Ultrashort-specific immunotherapy successfully treats seasonal allergic rhinoconjunctivitis to grass pollen.
Specific immunotherapy is a well-established treatment for allergic rhinoconjunctivitis; conventional regimens are lengthy, however, reducing convenience and cost-effectiveness. This study evaluated the efficacy and safety of an ultrashort course (four doses) of the immunotherapy Grass Modified Allergen Tyrosine Adsorbate (Allergy Therapeutics, Worthing, U.K.) monophosphoryl lipid A (MATA MPL). Subjects were randomized to receive four injections of either Grass MATA MPL (n = 514; 300-2000 standardized units/injection) or placebo (n = 514) before the grass pollen season. They used electronic diaries to record allergy symptoms and medication use during the pollen season. The primary end point was the difference between the mean combined symptom and medication scores in the Grass MATA MPL and placebo groups during the 4 local peak pollen weeks. The injection course was completed by 95.3 and 97.7% of the Grass MATA MPL and placebo groups, respectively, and was well tolerated. Grass MATA MPL treatment afforded a 13.4% benefit over placebo in the 4 peak pollen weeks (p = 0.0038). The benefit in subjects with 28 complete diary entries during the 4 peak pollen weeks was 26.9% (p = 0.0031). Significant benefits over placebo were observed in subjects with severe symptoms (17.1%; p = 0.0023), in those who had a history of allergic rhinoconjunctivitis for up to 35 years (up to 37.2%; p = 0.0059) and at sites with a higher burden of disease (38.3%; p < 0.0001). The ultrashort course of Grass MATA MPL was well tolerated and provided a significant benefit over placebo in relieving allergy symptoms.


High concentrations of aldehydes slow the reaction of cytoplasmic aldehyde dehydrogenase with thiol-group modifiers.
High concentrations of aldehydes slow the inactivation of cytoplasmic aldehyde dehydrogenase by disulfiram and also slow the reaction of the enzyme with 2,2'-dithiodipyridine. It is concluded that a low-affinity aldehyde-binding site is probably the site at which thiol-group modifiers react with aldehyde dehydrogenase, as well as being the active site for hydrolysis of 4-nitrophenyl acetate.


Streptococcus caledonicus sp. nov., isolated from sheep.
Five strains of an unidentified Gram-positive, catalase-negative, chain-forming coccus-shaped organism recovered from sheep in Scotland were characterized using phenotypic and molecular taxonomic methods. Based on morphological and biochemical criteria, the strains were tentatively identified as streptococci but they did not appear to correspond to any recognised species of the genus. Comparative 16S rRNA gene sequencing showed the strains were highly related to each other and confirmed their placement in the genus Streptococcus, with a maximum nucleotide identity of around 97 % to extant species. Best matches were with Streptococcus hillyeri followed by Streptococcus porci. Average nucleotide identity and in silico DNA-DNA hybridization values determined from whole-genome sequence were also consistent with the group representing a novel species. Best matches, again seen to S. hillyeri, followed by S. porci and S. plurextorum, were below accepted cut-off values for species delineation. Based on biochemical criteria and molecular genetic evidence, it is proposed that the unknown isolates from sheep be assigned to a new species of the genus Streptococcus as Streptococcus caledonicus sp. nov. The type strain of Streptococcus caledonicus is S784/96/1T=CCUG 73951T=NCTC 14363T.


Azadirachtin Attenuates Lipopolysaccharide-Induced ROS Production, DNA Damage, and Apoptosis by Regulating JNK/Akt and AMPK/mTOR-Dependent Pathways in Rin-5F Pancreatic Beta Cells.
Pancreatic inflammation and the resulting cellular responses have been implicated in pancreatitis, diabetes, and pancreatic cancer. Inflammatory responses due to the bacterial endotoxin, lipopolysaccharide (LPS), have been demonstrated to alter cellular metabolism, autophagy, apoptosis, and cell proliferation in different cell populations, and hence increases the risks for organ toxicity including cancer. The exact molecular mechanism is however not clear. In the present study, we investigated the role and mechanism of an antioxidant, azadirachtin (AZD), a limonoid extracted from the neem tree (Azadirachta indica), against LPS-induced oxidative stress in the pancreatic β-cell line, Rin-5F. We demonstrated that cells treated with LPS (1 µg/mL for 24 h) showed increased reactive oxygen species (ROS) production, DNA damage, cell cycle arrest, and apoptosis. Our results also showed that LPS induced alterations in the AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathways, suppressing autophagy and augmenting apoptosis. Treatment with Azadirachtin (25 µM for 24 h), on the other hand, rendered some degree of protection to the pancreatic cells from apoptosis by inducing the autophagy signals required for cell survival. These results may have significance in elucidating the mechanisms of pancreatic β-cell survival and death by balancing the molecular communication between autophagy and apoptosis under inflammatory and pathological conditions.


Genomic and Molecular Profiling of Human Papillomavirus Associated Head and Neck Squamous Cell Carcinoma Treated with Immune Checkpoint Blockade Compared to Survival Outcomes.
Recurrent/metastatic (R/M) head and neck squamous cell carcinoma (HNSCC) patients overall have a poor prognosis. However, human papillomavirus (HPV)-associated R/M oropharyngeal squamous cell carcinoma (OPSCC) is associated with a better prognosis compared to HPV-negative disease. Immune checkpoint blockade (ICB) is the standard of care for R/M HNSCC. However, whether HPV and its surrogate marker, p16, portend an improved response to ICB remains controversial. We queried the Caris Life Sciences CODEai database for p16+ and p16- HNSCC patients using p16 as a surrogate for HPV. A total of 2905 HNSCC (OPSCC, n = 948) cases were identified. Of those tested for both HPV directly and p16, 32% (251/791) were p16+ and 28% (91/326) were HPV+. The most common mutation in the OPSCC cohort was TP53 (33%), followed by PIK3CA (17%) and KMT2D (10.6%). TP53 mutations were more common in p16- (49%) versus the p16+ group (10%, p < 0.0005). Real-world overall survival (rwOS) was longer in p16+ compared to p16- OPSCC patients, 33.3 vs. 19.1 months (HR = 0.597, p = 0.001), as well as non-oropharyngeal (non-OP) HNSCC patients (34 vs. 17 months, HR 0.551, p = 0.0001). There was no difference in the time on treatment (TOT) (4.2 vs. 2.8 months, HR 0.796, p = 0.221) in ICB-treated p16+ vs. p16- OPSCC groups. However, p16+ non-OP HNSCC patients treated with ICB had higher TOT compared to the p16- group (4.3 vs. 3.3 months, HR 0.632, p = 0.016), suggesting that p16 may be used as a prognostic biomarker in non-OP HNSCC, and further investigation through prospective clinical trials is warranted.


[The experience of applying techniques of molecular genetics in identification of clinical strains Pseudomonas aeruginosa].
The article presents comparative analysis of application of common bacteriologic and molecular techniques to identify P. aerugenosa. The genetic detection was applied using polymerase chain reaction with genus-specific and species-specific primers and amplification of conservative sites of gens 16S pRNA with successive identification of bacteria by sequenation. It is established that 95% (151) of strains correspond to species of P. aerugenosa detected in primary bacteriologic laboratories and only 8 strains were not blue pus bacillus. Most of strains were closely congenial species: 2 isolates belonged to Pseudomonas aeruginosa group, 3 isolates to Pseudomonas putida group, 2 strains to Comamonas species and 1 isolate to Stenotrophomonas maltophilia species. The effectiveness of cultural technique of laboratory diagnostic was demonstrated concerning infections conditioned by P. aerugenosa. This conclusion does not eliminate application of molecular genetic trechnologies in complicated arbitral cases of bacteriologic analysis during monitoring of nosocomial infections.


Chitosan/poly(vinyl alcohol) hydrogel combined with Ad-hTGF-β1 transfected mesenchymal stem cells to repair rabbit articular cartilage defects.
The aim of this work is to explore the feasibility and therapeutic effect of repairing rabbit articular cartilage defects using thermo-sensitive chitosan/poly (vinyl alcohol) composite hydrogel engineered Ad-hTGF-β1-transfected bone marrow mesenchymal stem cells. Rabbit's bone marrow stromal cells (BMSCs) were obtained and cultured in vitro and transfected with a well-constructed Ad-hTGF-β1 vector, the cartilage phenotype of the transfected cells was tested by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Twenty-four New Zealand white rabbits with articular cartilage defects were randomly divided into four groups: group A was treated with CS/PVA gel and transfected BMSCs; group B received CS/PVA gel and un-transfected BMSCs; group C was treated with CS/PVA gel alone and group D was the untreated control group. Experimental animals of each group were killed at 16 weeks after operation. General observation, Masson's trichrome staining and collagen II immunohistological staining of the specimens were performed to evaluate the repair effect. The Wakitani scoring method was used to evaluate the repair effect. RT-PCR and Western blot confirmed that the hTGF-β1 gene was expressed in BMSCs and triggered the expression of specific markers of cartilage differentiation such as aggrecan mRNA and Collagen II in BMSCs after transfection with Ad-hTGF-β1. Sixteen weeks after operation, the defects in group A had smooth and flat surfaces, and the defects appeared to have completely healed, exhibiting almost the same color and texture as the surrounding cartilage. Masson's trichrome staining showed that the cell arrangement and density of regenerated cartilage tissue in group A was not significantly different from that of normal cartilage tissue. The immunohistochemical staining of Col II showed a strong expression in group A and weak expression in group B, but no expression in groups C and D. According to the Wakitani score, the difference between experimental group A and all of the other groups was statistically significant (P < 0.01). To conclude, as a thermosensitive and injectable scaffold material, CS/PVA gel engineered with BMSCs transfected with hTGF-β1 can effectively repair rabbit articular cartilage defects.


Osmotic cell swelling-induced ATP release mediates the activation of extracellular signal-regulated protein kinase (Erk)-1/2 but not the activation of osmo-sensitive anion channels.
Human intestine 407 cells respond to hypo-osmotic stress by the rapid release of ATP into the extracellular medium. A difference in the time course of activation as well as in the sensitivity to cytochalasin B treatment and BAPTA-AM [1,2-bis-(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid acetoxymethyl ester] loading suggests that ATP leaves the cell through a pathway distinct from volume-regulated anion channels. To evaluate a putative role for nucleotides as autocrinic/paracrinic factors in osmotic signalling, the effects of extracellular ATP on the regulation of volume-sensitive anion channels as well as on the hypotonicity-induced activation of extracellular signal-regulated protein kinases (Erk-1/2) were investigated. Micromolar concentrations of ATP were unable to elicit an isotope efflux from (125)I(-)-loaded cells by itself, but strongly potentiated the hypotonicity-provoked anion efflux through a Ca(2+)-dependent mechanism. The order of potency of nucleotides (ATP = UTP = ATP[S] > ADP = AMP >> adenosine = cAMP) indicated the involvement of P2Y(2) receptors. In contrast, millimolar concentrations of ATP markedly inhibited both the osmotically induced isotope efflux and whole-cell Cl(-) currents. Inhibition of whole-cell Cl(-) currents, not only by millimolar ATP but also by the purinoceptor antagonists suramin and reactive blue, was observed most prominently at depolarizing holding potentials, suggesting a direct interaction with volume-sensitive Cl(-) channels rather than interaction with purinoceptors. Both ATP and UTP, at submicromolar levels, were found to act as potent activators of Erk-1/2 in intestine 407 cells. Addition of the ATP hydrolase apyrase to the bath greatly reduced the hypotonicity-induced Erk-1/2 activation, but did not affect the swelling-induced isotope efflux or whole-cell Cl(-) currents. Furthermore, pre-treatment with suramin or reactive blue almost completely prevented the hypo-osmotic activation of Erk-1/2. The results indicate that extracellularly released ATP functions as an autocrinic/paracrinic factor that mediates hypotonicity-induced Erk-1/2 activation but does not serve as an activator of volume-sensitive compensatory Cl(-) currents.


3-[4-(Dimethyl-amino)-phen-yl]-1-(4a,8-dimethyl-1,2,3,4,4a,5,6,8a-octa-hydro-naphthalen-2-yl)prop-2-en-1-one.
The title compound, C(23)H(31)NO, was semisynthesized from isocostic acid, isolated from the aerial part of Inula Viscosa- (L) Aiton [or Dittrichia Viscosa- (L) Greuter]. The cyclo-hexene ring has a half-chair conformation, whereas the cyclo-hexane ring displays a chair conformation. The dihedral angle between the latter ring and its substituent is 83.6 (7)°.


Does inpatient palliative care consultation impact outcomes following hospital discharge? A narrative systematic review.
While most patients desire to die at home or in a community-based hospice, the transition from hospital to community settings often lacks streamlined coordination of care to ensure that adequate support is provided in the preferred care setting. The impact of hospital-based palliative care consultations on post-discharge care and outcomes has not been extensively studied. The aim of this study was to appraise available research on the impact of inpatient palliative care consultations on transitions from hospital to community settings. We conducted a narrative systematic review and used the Effective Public Health Practice Project tool to appraise the quality of selected studies. Studies were included if they assessed the transition from hospital to community and examined outcomes after an inpatient palliative care consultation. A protocol for this study was registered and published in PROSPERO, Centre for Reviews and Dissemination (ID: CRD42018094924). We searched for quantitative studies indexed in PubMED, CINAHL and Cochrane and published between 1 January 1 2000 and 11 March 2018. Our search retrieved 2749 articles. From these, 123 articles were full-text screened and 15 studies met our inclusion criteria. Studies reported that inpatient palliative care consultations are associated with high rates of discharge to community settings, greater provision of services post-discharge, improved coordination and lower rates of rehospitalization. Existing evidence suggest that inpatient palliative care consultations have a positive impact on patient outcomes and transitions to the community, demonstrating the potential to improve patient quality of life and relieve overburdened acute care systems.


Fruit and vegetable pricing by demographic factors in the Birmingham, Alabama, metropolitan area, 2004-2005.
Fruit and vegetable cost may influence consumption. Because the contextual environment influences food outlet type and availability, we wanted to determine whether neighborhood demographics were associated with prices of fruits and vegetables. We surveyed 44 grocery stores in the Birmingham, Alabama, metropolitan area to determine prices of 20 fruits and vegetables. Stores were geocoded and linked to the corresponding Census 2000 block group to obtain data for the independent variables - percentage African American, percentage with at least a high school diploma, and percentage of households below the poverty level. We conducted multiple linear regressions to estimate these predictors for each fruit and vegetable's mean price per serving during 2 seasons (fall/winter 2004, spring/summer 2005). In the fall, we found no significant relationships between the predictors and prices of any fruits and vegetables in the survey. In the spring, the percentage who had at least a high school diploma was a predictor of price per serving for potatoes (beta = 0.001, P = .046). Neighborhood demographics have little consistent influence on fruit and vegetable prices in Birmingham, Alabama, which may be a function of grocery store density, transportation patterns, and shopping patterns. The regional setting of the food environment has implications for food availability, variety, and price.


Structural studies on phenylalanine hydroxylase and implications toward understanding and treating phenylketonuria.
Mutations in the gene encoding for phenylalanine hydroxylase (PAH) result in phenylketonuria (PKU) or hyperphenylalaninemia (HPA). Several 3-dimensional structures of truncated forms of PAH have been determined in our laboratory and by others, using x-ray crystallographic techniques. These structures have allowed for a detailed mapping of the >250 missense mutations known to cause PKU or HPA found throughout the 3 domains of PAH. This structural information has helped formulate rules that might aid in predicting the likely effects of unclassified or newly discovered PAH mutations. Also, with the aid of recent crystal structure determinations of co-factor and substrate analogs bound at the PAH active site, the recently discovered tetrahydrobiopterin-responsive PKU/HPA genotypes can be mapped onto the PAH structure, providing a molecular basis for this tetrahydrobiopterin response.


Sports hernias.
Sportsman's hernia is an increasingly recognized cause of chronic groin pain in athletes. Although the definition is controversial, it is a condition of chronic inguinal/pubic exertional pain caused by rectus abdominal wall weakness or injury without a palpable hernia, usually affecting high-performance male athletes. Diagnosis is made after careful history and physical examination. Some radiographic studies such as ultrasound or MRI may be helpful in evaluating these patients and ruling out other pathology, although no radiographic study can rule out sportsman's hernias. Because sports hernias are not true hernias but an injury in the rectus insertion, unilateral or bilateral rectus reattachment is the most appropriate surgical treatment. This reattachment may be done in combination with adductor release in the setting of adductor pain or weakness on physical examination. Other surgical repairs (eg, Lichtenstein, Shouldice, Kugel, laparoscopic) do not stabilize the pelvis and tend not to be as successful. In the motivated patient, after surgical repair and physical rehabilitation, 95% are free of pain and able to return to competitive sports.


The role of the high-density lipoprotein receptor SR-BI in cholesterol metabolism.
The HDL receptor scavenger receptor class B type I (SR-BI), which mediates selective HDL cholesterol uptake, plays a role in murine HDL metabolism, reverse cholesterol transport and whole-body cholesterol homeostasis. SR-BI is found in the liver, where its expression is regulated by estrogen, dietary cholesterol and fat, and controls murine plasma HDL cholesterol levels and bile cholesterol secretion. SR-BI is also highly expressed in rodent steroidogenic cells, where it facilitates cholesterol uptake for storage or steroid hormone synthesis and where its expression is regulated by trophic hormones. The detailed mechanism(s) underlying SR-BI-mediated selective cholesterol uptake have not yet been elucidated. Further analysis of the molecular and cellular bases of SR-BI regulation and function should provide new insights into the physiology and pathophysiology of cholesterol metabolism.


Carbenoxolone and gastric Technetium-99m pertechnetate uptake in patients with gastric ulceration.
Technetium-99m pertechnetate is known to concentrate in the stomach following intravenous injection, but there is controversy on the cellular site of this uptake, with evidence for both the mucous and parietal cells being primarily responsible. Gastric uptake following intravenous pertechnetate has been studied in patients with non-ulcer dyspepsia and peptic ulcer, using a gamma camera and computer processing with regions of interest. Under identical conditions in any one patient, quantification of uptake at 20 min is reproducible to an accuracy of +/- 6%. Six patients with gastric ulcer were studied before and 3 weeks after starting treatment with carbenoxolone. There was a rise in uptake in 5 subjects with a mean percentage increase of 25%. The effect of cimetidine was studied in six patients with either gastric or duodenal ulcer. There was little change in uptake (mean 6%). These preliminary data would support the hypothesis that concentration of pertechnetate occurs primarily in the mucous cell.


B-cell adaptor for PI3K (BCAP) negatively regulates Toll-like receptor signaling through activation of PI3K.
Toll-like receptors (TLRs) recognize pathogens and their components, thereby initiating immune responses to infectious organisms. TLR ligation leads to the activation of NF-κB and MAPKs through well-defined pathways, but it has remained unclear how TLR signaling activates PI3K, which provides an inhibitory pathway limiting TLR responses. Here, we show that the signaling adapter B-cell adaptor for PI3K (BCAP) links TLR signaling to PI3K activation. BCAP-deficient macrophages and mice are hyperresponsive to TLR agonists and have reduced PI3K activation. The ability of BCAP to inhibit TLR responses requires its capacity to bind PI3K. BCAP is constitutively phosphorylated and associated with the p85 subunit of PI3K in macrophages. This tyrosine-phosphorylated BCAP is transiently enriched in the membrane fraction in response to LPS treatment, suggesting a model whereby TLR signaling causes the phosphorylation of the small amount of BCAP that is associated with membranes in the resting state or the translocation of phosphorylated BCAP from the cytoplasm to the membrane. This accumulation of tyrosine-phosphorylated BCAP at the membrane with its associated PI3K would then allow for the catalysis of Ptd Ins P2 to Ptd Ins P3 and downstream PI3K-dependent signals. Therefore, BCAP is an essential activator of the PI3K pathway downstream of TLR signaling, providing a brake to limit potentially pathogenic excessive TLR responses.


FPF1 promotes flowering in Arabidopsis.
We have characterized the gene flowering promoting factor1 (FPF1), which is expressed in apical meristems immediately after the photoperiodic induction of flowering in the long-day plants mustard and Arabidopsis. In early transition stages, expression is only detectable in the peripheral zone of apical meristems; however, later on, it can also be found in floral meristems and in axillary meristems that form secondary inflorescences. The FPF1 gene encodes a 12.6-kD protein that has no homology to any previously identified protein of known function. Constitutive expression of the gene in Arabidopsis under control of the cauliflower mosaic virus 35S promoter resulted in a dominant heritable trait of early flowering under both short- and long-day conditions. Treatments with gibberellin (GA) and paclobutrazol, a GA biosynthesis inhibitor, as well as crosses with GA-deficient mutants indicate that FPF1 is involved in a GA-dependent signaling pathway and modulates a GA response in apical meristems during the transition to flowering.


Zinc Supplementation During Pregnancy Alleviates Lipopolysaccharide-Induced Glial Activation and Inflammatory Markers Expression in a Rat Model of Maternal Immune Activation.
Maternal immune activation (MIA) model has been profoundly described as a suitable approach to study the pathophysiological mechanisms of neuropsychiatric disorders, including schizophrenia. Our previous study revealed that prenatal exposure to lipopolysaccharide (LPS) induced working memory impairments in only male offspring. Based on the putative role of prefrontal cortex (PFC) in working memory process, the current study was conducted to examine the long-lasting effect of LPS-induced MIA on several neuroinflammatory mediators in the PFC of adult male pups. We also investigated whether maternal zinc supplementation can alleviate LPS-induced alterations in this region. Pregnant rats received intraperitoneal injections of either LPS (0.5 mg/kg) or saline on gestation days 15/16 and supplemented with ZnSO4 (30 mg/kg) throughout pregnancy. At postnatal day 60, the density of both microglia and astrocyte cells and the expression levels of IL-6, IL-1β, iNOS, TNF-α, NF-κB, and GFAP were evaluated in the PFC of male pups. Although maternal LPS treatment increased microglia and astrocyte density, number of neurons in the PFC of adult offspring remained unchanged. These findings were accompanied by the exacerbated mRNA levels of IL-6, IL-1β, iNOS, TNF-α, NF-κB, and GFAP as well. Conversely, prenatal zinc supplementation alleviated the mentioned alterations induced by LPS. These findings support the idea that the deleterious effects of prenatal LPS exposure could be attenuated by zinc supplementation during pregnancy. It is of interest to suggest early therapeutic intervention as a valuable approach to prevent neurodevelopmental deficits, following maternal infection. Schematic diagram describing the experimental timeline. On gestation days (GD) 15 and 16, pregnant dams were administered with intraperitoneal injections of either LPS (0.5 mg/kg) or vehicle and supplemented with ZnSO4 (30 mg/kg) throughout pregnancy by gavage. The resulting offspring were submitted to qPCR, immunostaining, and morphological analysis at PND 60. Maternal zinc supplementation alleviated increased expression levels of inflammatory mediators and microglia and astrocyte density induced by LPS in the PFC of treated offspring. PND postnatal day, PFC prefrontal cortex.


Carbon dots derived from kanamycin sulfate with antibacterial activity and selectivity for Cr6+ detection.
Among antibacterial nanomaterials, carbon dots (CDs) have attracted much attention because of their unique physical and chemical properties and good biosafety. In this study, kanamycin sulfate (Kan), a broad-spectrum antibiotic, was used to synthesize novel carbon dots (CDs-Kan) by a one-step hydrothermal method. CDs-Kan showed good inhibitory effects on Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus. Further, scanning electron microscopy revealed that treatment with CDs-Kan and Kan resulted in the same phenomena. In particular, the morphologies of S. aureus cells treated with CDs-Kan and Kan became smaller and irregular, whereas the surfaces of E. coli cells protruded and formed vesicles. These results indicated that CDs-Kan was shown to retain the good antibacterial activity of Kan as well as its main bactericidal functional groups, namely, the amino sugar and amino cyclic alcohol, We refer to this phenomenon as the "preservation property". We also found that CDs-Kan has good biocompatibility and nontoxic properties. Moreover, CDs-Kan was successfully applied to the biological imaging of fungi and plant cells. In addition, CDs-Kan could be used as a fluorescent probe for the quick, sensitive, and selective detection of Cr6+. Therefore, CDs-Kan not only retained the good bacteriostatic properties of Kan but also expanded its application in bioimaging and biosensors.


Joint source-channel coding for wireless object-based video communications utilizing data hiding.
In recent years, joint source-channel coding for multimedia communications has gained increased popularity. However, very limited work has been conducted to address the problem of joint source-channel coding for object-based video. In this paper, we propose a data hiding scheme that improves the error resilience of object-based video by adaptively embedding the shape and motion information into the texture data. Within a rate-distortion theoretical framework, the source coding, channel coding, data embedding, and decoder error concealment are jointly optimized based on knowledge of the transmission channel conditions. Our goal is to achieve the best video quality as expressed by the minimum total expected distortion. The optimization problem is solved using Lagrangian relaxation and dynamic programming. The performance of the proposed scheme is tested using simulations of a Rayleigh-fading wireless channel, and the algorithm is implemented based on the MPEG-4 verification model. Experimental results indicate that the proposed hybrid source-channel coding scheme significantly outperforms methods without data hiding or unequal error protection.


Clear cell renal cell carcinoma with wild-type von Hippel-Lindau gene: a non-existent or new tumour entity?
The current World Health Organisation (WHO) classification of renal tumours is based on characteristic histological features or specific molecular alterations. von Hippel-Lindau (VHL) alteration is the hallmark of clear cell renal cell carcinoma (RCC). After identification of the MiT translocation family of tumours, clear cell papillary renal cancer and others, the group of ccRCC with wild-type VHL is small. TCEB1 mutation combined with chromosome 8q loss is an emerging tumour entity with wild-type VHL. Inactivation of TCEB1 increases HIF stabilisation via the same mechanism as VHL inactivation. Importantly, recent molecular analyses suggest the existence of another 'VHL wild-type' evolutionary subtype of clear cell RCC in addition to TCEB1 mutated RCC and clear cell papillary renal cancer. These tumours are characterised by an aggressive behaviour, high tumour cell proliferation rate, elevated chromosomal instability and frequent presence of sarcomatoid differentiation. Future clinicopathological studies will have to provide data to determine whether TCEB1 tumours and clear cell RCC with wild-type VHL are separate tumour entities or represent variants of a clear cell RCC tumour family.


The psychiatrist's involvement with aged patients.
The author studied via questionnaire psychiatrists' involvement with older patients in relation to their own ages and the nature of their practices. Older psychiatrists see a higher proportion of older patients, advocate more comprehensive treatment, and find older patients more interesting and gratifying. Psychiatrists in general psychiatry and those who have a primarily inpatient practice see the largest proportion of older patients.


Nitrogen balance in patients receiving either fat or carbohydrate for total intravenous nutrition.
Eighteen pre and/or postoperative patients underwent TIN using either fat or carbohydrate as a caloric substrate source for a similar period. Positive nitrogen balance was achieved with either solution for an equal number of days when balance studies were complete. Both groups demonstrated weight gain but it was more marked in the carbohydrate TIN group. One patient in the carbohydrate TIN group was changed to Intralipid because of a catheter related complication. This patient continued to show positive nitrogen balance while on Intralipid. There were 4 deaths in the 18 patients unrelated to TIN. It is concluded that Intralipid, when given through a peripheral vein with a nitrogen source, can produce positive nitrogen balance and is a safe and effective means for doing this.


Keratinocyte-derived IL-24 plays a role in the positive feedback regulation of epidermal inflammation in response to environmental and endogenous toxic stressors.
Keratinocytes are the major cellular components of human epidermis and play a key role in the modulating cutaneous inflammation and toxic responses. In human chronic skin diseases, the common skin inflammatory phenotypes like skin barrier disruption and epidermal hyperplasia are manifested in epidermal keratinocytes by interactions with T helper (Th) cells. To find a common gene expression signature of human keratinocytes in chronic skin diseases, we performed a whole genome microarray analysis on normal human epidermal keratinocytes (NHKs) treated with IFNγ, IL-4, IL-17A or IL-22, major cytokines from Th1, Th2, Th17 or Th22 cells, respectively. The microarray results showed that the four genes, IL-24, PDZK1IP1, H19 and filaggrin, had common expression profiles in NHKs exposed to Th cell cytokines. In addition, the acute phase pro-inflammatory cytokines, IL-1β, IL-6 and TNFα, also change the gene transcriptional profile of IL-24, PDZK1IP1, H19, and filaggrin in NHKs as those of Th cytokines. Therefore, the signature gene set, consisting of IL-24, PDZK1IP1, H19, and filaggrin, provides essential insights for understanding the process of cutaneous inflammation and toxic responses. We demonstrate that environmental toxic stressors, such as chemical irritants and ultraviolet irradiation stimulate the production of IL-24 in NHKs. IL-24 stimulates the JAK1-STAT3 and MAPK pathways in NHKs, and promotes the secretion of pro-inflammatory mediators IL-8, PGE2, and MMP-1. These results suggest that keratinocyte-derived IL-24 participates in the positive feedback regulation of epidermal inflammation in response to both endogenous and environmental toxic stressors.


Altered balance between matrix metalloproteinases and their inhibitors in experimental biliary fibrosis.
A rat model of common bile duct ligation (BDL)-induced hepatic fibrosis was used to assess the expression and activities of collagen-degrading proteinases and their inhibitors during the progression of fibrosis. Expression of four members of the matrix metalloproteinase (MMP) family (MMP-2/gelatinase A, MMP-3, MMP-9/gelatinase B, and MMP-13) and three tissue inhibitors of metalloproteinases-1, -2, and -3 (TIMP-1, TIMP-2, and TIMP-3) were evaluated by Northern blot analysis of RNA from liver tissue isolated at 0, 2, 5, 10, 20, and 30 days after either a BDL or sham operation. In addition, we analyzed free gelatinase and TIMP activities by zymography and reverse zymography, respectively. We found that the proteolytic activities of MMP-2 and MMP-9 increased by 2 days after ligation, reached maximal levels at day 10, and remained high through the study period, whereas the gelatinolytic activities in plasma were unchanged. The increase in gelatinase activities was accompanied by an increase in the TIMP mRNA transcripts. TIMP-1 transcripts appeared at day 2, increased until day 10, and remained elevated throughout the study period. TIMP-2 and TIMP-3 transcripts become detectable on day 10 and remained stable afterwards. No corresponding increase in TIMP protein activity was detected by reverse zymography. This appears to result from the formation of TIMP/MMP complexes. These findings indicate a likely surplus in the BDL model of fibrosis of free gelatinases as compared with the TIMPs. Thus, excessive TIMP production is not a sufficient explanation for the observed extracellular matrix accumulation, but complex changes in the local MMP/TIMP balance may underlie the pathomechanisms of fibrosis.