In-line sample trap columns with diatomite for large-volume injection in CZE-IM-MS.
The analysis of low-abundant compounds with capillary zone electrophoresis-drift-tube ion mobility spectrometry-mass spectrometry (CZE-DTIMS-MS) is compromised due to the low injectable sample volumes in CZE and low duty cycle in DTIMS. Fritless packed in-line trap columns, using solid-phase extraction sorbent particles, have been used to increase injection volumes in CZE, but these columns are difficult to prepare and exhibit rapidly increasing back pressures. To provide smooth and complete filling of trap columns as well as to ensure higher and sustained flow rates though the columns, blends of cation and anion exchange particles with diatomite were used. The application of diatomite blends ensured the use of trap columns for at least 100 injections, with maximum injection volumes over 10 µl, which corresponds to an enrichment factor of more than 1000 over conventional injections in CZE-MS, enabling the detection of low nM concentrations of N-glycans with CZE-IMS-MS.


Inhibitory effect of Newtonia extracts and myricetin-3-o-rhamnoside (myricitrin) on bacterial biofilm formation.
Diarrhoea is a major health issue in both humans and animals and may be caused by bacterial, viral and fungal infections. Previous studies highlighted excellent activity of Newtonia buchananii and N. hildebrandtii leaf extracts against bacterial and fungal organisms related to diarrhoea-causing pathogens. The aim of this study was to isolate the compound(s) responsible for antimicrobial activity and to investigate efficacy of the extracts and purified compound against bacterial biofilms. The acetone extract of N. buchananii leaf powder was separated by solvent-solvent partitioning into eight fractions, followed by bioassay-guided fractionation for isolation of antimicrobial compounds. Antibacterial activity testing was performed using a broth microdilution assay. The cytotoxicity was evaluated against Vero cells using a colorimetric MTT assay. A crystal violet method was employed to test the inhibitory effect of acetone, methanol: dichloromethane and water (cold and hot) extracts of N. buchananii and N. hildebrandtii leaves and the purified compound on biofilm formation of Pseudomonas aeruginosa, Escherichia coli, Salmonella Typhimurium, Enterococcus faecalis, Staphylococcus aureus and Bacillus cereus. Myricetin-3-o-rhamnoside (myricitrin) was isolated for the first time from N. buchananii. Myricitrin was active against B. cereus, E. coli and S. aureus (MIC = 62.5 μg/ml in all cases). Additionally, myricitrin had relatively low cytotoxicity with IC50 = 104 μg/ml. Extracts of both plant species had stronger biofilm inhibitory activity against Gram-positive than Gram-negative bacteria. The most sensitive bacterial strains were E. faecalis and S. aureus. The cold and hot water leaf extracts of N. buchananii had antibacterial activity and were relatively non-cytotoxic with selectivity index values of 1.98-11.44. The purified compound, myricitrin, contributed to the activity of N. buchananii but it is likely that synergistic effects play a role in the antibacterial and antibiofilm efficacy of the plant extract. The cold and hot water leaf extracts of N. buchananii may be developed as potential antibacterial and antibiofilm agents in the natural treatment of gastrointestinal disorders including diarrhoea in both human and veterinary medicine.


The effects of D-penicillamine on a murine model of oxygen-induced retinopathy.
To determine the effect of intraperitoneal and intravitreal D-penicillamine (DPA) on retinal neovascularization in a murine model of oxygen-induced retinopathy. On postnatal day 7, 16 mice were injected intraperitoneally with 300 mg/kg/day DPA for 3 days followed by 50 mg/kg/day for 7 days. A different group of 7 mice were injected intraperitoneally with 600 mg/kg/day DPA for 3 days followed by 100 mg/kg/day for 7 days. A third group of 14 mice were injected with 1,500 mg/kg/day DPA for 2 days; a control cohort of 17 mice received intraperitoneal phosphate-buffered saline (PBS). An additional 15 mice underwent intravitreal injection of 1 μL of 100 mg/mL DPA in the right eye and 1 μL PBS intravitreally in the left eye as a control. All groups were placed in a 75% oxygen chamber for 7 days then room air for 3 days before being sacrificed and enucleated. The retinas were stained and flat-mounted to determine the severity of retinal neovascularization by quantifying neovascular buds. After intraperitoneal injection, the mean number of glomeruli and tubules was similar in the DPA and PBS groups (P = 1.0), regardless of DPA dosage. The dosage of 1,500 mg/kg/day proved to be uniformly lethal. After intravitreal injections, the mean number of glomeruli (P = 0.16) and tubules (P = 0.7) were similar in the DPA and PBS groups. Neither intraperitoneal nor intravitreal injection of DPA inhibits retinal neovascularization in a murine model of oxygen-induced retinopathy.


In vivo insulin antagonism but evanescent in vitro tissue effect in rats with growth hormone-secreting tumors.
Rats bearing mammosomatotropic tumors have raised insulin but lowered glucose concentrations. To determine if growth hormone (GH) secreted by these tumors causes insulin antagonism, pancreatic suppression tests utilizing infusions (per kg per min) of glucose (8 mg), insulin (200 ng) and somatostatin (1.4 micrograms) for 130 min were performed. Although the steady state plasma glucose and insulin levels (mean of 90, 100, 110, 120 and 130 min samples) were similar in 8 control and 13 tumor-bearing rats, the decrease from the already depressed basal glucose concentration (mmoles/l +/- SE) in the tumor animals was less than in the controls (0.90 +/- 0.30 vs. 2.56 +/- 0.040, p less than 0.005). Since the interpretation of these results was not entirely clear, glucose and insulin-glucose tolerance tests were performed. The glucose disappearance rates (%/min +/- SE) in the glucose tolerance test were lower in 17 tumor rats (2.00 +/- 0.13) compared to 17 control animals (2.51 +/- 0.22). This difference just missed statistical significance (t = 2.00, value of 2.04 necessary for p = 0.05). The decrease occurred in the presence of increased insulin (nmoles/l X 16 min) levels (4.29 +/- 0.38 vs. 2.58 +/- 0.29, p less than 0.005) suggesting insulin antagonism. The glucose disappearance rates (%/min +/- SE) in the insulin-glucose tolerance test were less in 12 tumor-bearing rats compared to 11 control animals (2.80 +/- 0.29 vs. 4.12 +/- 0.35, p less than 0.02). Thus, these GH-secreting tumors cause insulin antagonism in vivo. Freshly isolated hepatocytes from these tumor-bearing animals manifest decreased insulin binding and action (Diabetologia 25:60, 1983). In the present study, however, insulin binding and action (net glucose-C14 incorporation into glycogen) were normal after the hepatocytes were cultured for two days. This suggests that the changes induced by GH in vivo that lead to insulin antagonism are short-lived.


Propagation of the marine dinoflagellate Amyloodinium ocellatum under germ-free conditions.
Germ-free infections of Amyloodinium ocellatum were produced on both living fish and in organ cultures. Exposing gnotobiotic guppies (Poecilia reticulata) to 125 dinospores in multiwell tissue culture plates produced nonlethal infections that could be serially propagated. Exposure to 250 or more parasites killed the fish during the first infection cycle, but if the dead fish were incubated in a cell culture medium/seawater mixture, the parasites could survive and reproduce for up to 2 wk in these organ cultures. Organ cultures containing only seawater or those containing bacteria did not support the prolonged survival of Amyloodinium.


SCORE and REGICOR function charts underestimate the cardiovascular risk in Spanish patients with rheumatoid arthritis.
Our objective was to determine which one of the two function charts available in Spain to calculate cardiovascular (CV) risk, Systematic COronary Risk Evaluation (SCORE) or Framingham-REgistre GIroní del COR (REGICOR), should be used in patients with rheumatoid arthritis (RA). A series of RA patients seen over a one-year period without history of CV events were assessed. SCORE, REGICOR, modified (m)SCORE and mREGICOR according to the European League Against Rheumatism (EULAR) recommendations were applied. Carotid ultrasonography (US) was performed. Carotid intima-media thickness (cIMT) > 0.90 mm and/or carotid plaques were used as the gold standard test for severe subclinical atherosclerosis and high CV risk (US+). The area under the receiver operating curves (AUC) for the predicted risk for mSCORE and mREGICOR were calculated according to the presence of severe carotid US findings (US+). We included 370 patients (80% women; mean age 58.9 ± 13.7 years); 36% had disease duration of 10 years or more; rheumatoid factor (RF) and/or anticyclic citrullinated peptide (anti-CCP) were positive in 68%; and 17% had extra-articular manifestations. The EULAR multiplier factor was used in 122 (33%) of the patients. The mSCORE was 2.16 ± 2.49% and the mREGICOR 4.36 ± 3.46%. Regarding US results, 196 (53%) patients were US+. The AUC mSCORE was 0.798 (CI 95%: 0.752 to 0.844) and AUC mREGICOR 0.741 (95% CI; 0.691 to 0.792). However, mSCORE and mREGICOR failed to identify 88% and 91% of US+ patients. More than 50% of patients with mSCORE ≥1% or mREGICOR >1% were US+. Neither of these two function charts was useful in estimating CV risk in Spanish RA patients.


Silvernanoparticle-induced hemolysis confounded with direct antiglobulin test-negative autoimmune hemolytic anemias diagnosis.
We describe here the first patient with recurrent hemolysis related to disinfectant containing silver nanoparticles (AgNps). A 58-year-old chemist repeatedly experienced DAT-negative (Coombs-negative) hemolysis during the last 5 years. He was treated with a number of immunosuppressive drugs including 18 times rituximab. The attempt to treat him with cyclosporine A served only to increase the rate of hemolysis. Only by chance, we revealed that the patient regularly used a hand disinfectant containing AgNps. Serological testing was performed using standard techniques. Eryptosis was measured by binding annexin to exposed phosphatidylserine (PS) of the circulating red blood cells (RBCs). Antiglobulin tests remained negative, and PS exposing RBCs were detected two times during the last hemolytic episodes. Hemolysis completely disappeared following discontinuation of AgNp containing products. AgNps are increasingly being used in a large variety of products. Recently, it was reported that they induce in vitro prohemolytic and procoagulant effects via oxidative stress and eryptosis. The clinical findings imply the hemolysis was provoked by the patient's regular use of cleansing products containing AgNps. Our finding might help to explain the etiology of hemolytical disorders that may remain obscure in many cases.


Development of a self-report measure to assess sleep satisfaction: Protocol for the Suffolk Sleep Index (SuSI).
Good sleep is essential for health but there is no consensus on how to define and measure people's understanding of good sleep. To date, people's perceptions of a good night's sleep have been, almost exclusively, conceptualized under the lens of sleep quality, which refers to objective characteristics of good sleep, such as such as ease and time needed to fall asleep, hours of sleep, and physical symptoms during sleep and upon awakening. A related, yet different construct, sleep satisfaction, refers to perceived positive affect about one's sleep experience and has, to date, received little attention. This research focuses on sleep satisfaction, rather than sleep quality, and aims to develop a self-report measure to assess sleep satisfaction in an English adult population. As the measure will be developed in Suffolk, England, and its primary intended users are Suffolk community members, it is labelled the Suffolk Sleep Index (SuSI). The SuSI will draw from principles of community-based participatory research, following a seven-phase developmental process comprising literature review, interviews with Suffolk community members, synthesis of literature review and interview findings, pre-testing, pilot-testing, scale evaluation, and refinement. The present research will also investigate indices related to sleep satisfaction, including the community's general health status, lifestyle factors and socio-economic status. The research will add to the limited, yet emerging body of research on perceived sleep satisfaction and its measurement. To our knowledge, a valid and reliable sleep satisfaction self-report measure has not been developed in the United Kingdom previously.


The association of stress and work hours with sleep duration and insomnia symptoms among U.S.-born and Foreign-born Black adults.
To examine associations between stress and work hours with sleep duration and insomnia symptoms among U.S.-born and Foreign-born Black adults. A cross-sectional analysis of the National Health Interview Survey. Black adults (N = 16,315) with a mean age of 40 ± 0.1 years, mostly women (52.7%), college graduates (40.7%), and U.S.-born (82.5%). Participants self-reported sleep duration, insomnia symptoms, frequent stress (yes/no), and work hours in the prior week (1-39 hours, 40 hours, ≥41 hours). Poisson regression with robust variance was used to test associations overall and by nativity. Working ≥41 hours, frequent stress, and short sleep duration (<7 hours) were reported by 22%, 24.9%, and 43% of participants, respectively. U.S.-born and Foreign-born Black adults who reported frequent stress and working ≥41 hours vs. 40 hours had 60% and 19% higher prevalence of short sleep duration (adjusted prevalence ratio [aPR] = 1.60, 95% confidence interval [CI]: 1.47, 1.74 and aPR = 1.19, 95% CI: 1.12, 1.25, respectively). Foreign-born Black adults who worked 1-39 hours vs. 40 hours, had higher prevalence of short sleep duration, aPR = 1.18, 95% CI: 1.01, 1.37. U.S.-born Black adults who worked ≥41 hours vs. 40 hours had higher prevalence of insomnia symptoms (trouble falling asleep: aPR = 1.33 [95% CI: 1.13, 1.56], trouble staying asleep: aPR = 1.33 [95% CI: 1.16, 1.53]). Frequent stress and working ≥41 hours are likely salient determinants of sleep health for U.S. and Foreign-born Black individuals. Further, less work hours (Foreign-born) while longer work hours (U.S.-born) were associated with short sleep duration. Stress and work hours may be factors for sleep health interventions among Black adults.


Expression of NAD+ dependent 15-hydroxyprostaglandin dehydrogenase and protection of prostaglandins in human hair follicle.
NAD(+) dependent 15-hydroxyprostaglandin dehydrogenate (15-PGDH) catalyses oxidation of 15(S)-hydroxyl group of prostaglandins and as a result inactivates their physiological potential. Positive effects of prostaglandins or prostaglandin analogues were reported on terminal hair, vellus hair or eyelash growth and a complex prostaglandin network was recently described in human hair follicle. In the present study, we showed that 15-PGDH was expressed in human hair follicle mainly in melanocytes and keratinocytes, which brought us to consider this enzyme as a possible target to sustain local prostaglandin production. Using a recombinant enzymatic strategy, specific 15-PGDH inhibitors were screened. We identified a thiazolidine dione derivative exhibiting efficacy on follicular outer root sheath keratinocytes, since it concomitantly decreased the production of deactivated 13,14 dihydro 15-ketoprostaglandin F(2alpha) and sustained prostaglandin F(2alpha)in vitro production. In the context of recent interest in prostaglandins and prostaglandin analogues as hair regrowth agents, we postulated that the use of selected 15-PGDH inhibitors could reinforce or prolong the effect of these physiological mediators on hair and skin.


Reflections on the Applications of Immunohistochemistry to Understanding the Maintenance of Electrochemical Gradients in the Inner Ear.
The author of the accompanying classic paper from the Journal of Histochemistry and Cytochemistry (Crouch JJ, Sakaguchi N, Lytle C, Schulte BA. Immunohistochemical localization of the Na-K-Cl Co-transporter (NKCC1) in the Gerbil Inner Ear. Journal of Histochemistry & Cytochemistry 1997;45(6):773-778) comments on how the immunohistochemical techniques used in the study provided critical new information that helped define the cellular and molecular mechanisms involved in the generation and maintenance of electrochemical gradients in the ear, particularly the presence of the Na,K,2Cl symporter (NKCC) in the inner ear. (J Histochem Cytochem 71: 509-510, 2023).


Calculating evidence-based renal replacement therapy - Introducing an excel-based calculator to improve prescribing and delivery in renal replacement therapy - A before and after study.
Transferring the theoretical aspect of continuous renal replacement therapy to the bedside and delivering a given "dose" can be difficult. In research, the "dose" of renal replacement therapy is given as effluent flow rate in ml kg-1 h-1. Unfortunately, most machines require other information when they are initiating therapy, including blood flow rate, pre-blood pump flow rate, dialysate flow rate, etc. This can lead to confusion, resulting in patients receiving inappropriate doses of renal replacement therapy. Our aim was to design an excel calculator which would personalise patient's treatment, deliver an effective, evidence-based dose of renal replacement therapy without large variations in practice and prolong filter life. Our calculator prescribes a haemodialfiltration dose of 25 ml kg-1 h-1 whilst limiting the filtration fraction to 15%. We compared the episodes of renal replacement therapy received by a historical group of patients, by retrieving their data stored on the haemofiltration machines, to a group where the calculator was used. In the second group, the data were gathered prospectively. The median delivered dose reduced from 41.0 ml kg-1 h-1 to 26.8 ml kg-1 h-1 with reduced variability that was significantly closer to the aim of 25 ml kg-1.h-1 (p < 0.0001). The median treatment time increased from 8.5 h to 22.2 h (p = 0.00001). Our calculator significantly reduces variation in prescriptions of continuous veno-venous haemodiafiltration and provides an evidence-based dose. It is easy to use and provides personal care for patients whilst optimizing continuous veno-venous haemodiafiltration delivery and treatment times.


Increased WD40 motifs in Planctomycete bacteria and their evolutionary relevance.
Species of the family Planctomycetes have a complex intracellular structure, which is distinct from that of the majority of non-Planctomycetes bacteria. At present, genomic evidence of the evolution of intracellular complexity is lacking, cognitions of Planctomycetes's intracellular structure mainly rely on electron microscope observation. As the presence of WD40 motifs in eukaryotic proteins probably links to intracellular complexity, bioinformatic studies were conducted to detect and enumerate WD40 motifs, WD40 domains, and WD40 motif-bearing proteins in the genomes of 11 Planctomycetes species, 2775 non-Planctomycetes bacteria, and 63 representative eukaryotes. Compared to non-Planctomycetes bacteria (average 5 WD40 motifs and 1 WD40 motif-bearing protein per genome), a large increase in the number of WD40 motifs in Planctomycetes species (average 116 WD40 motifs and 26 WD40 motif-bearing proteins per genome) was observed. However, the average number of WD40 motifs in Planctomycetes species was significantly lower than that of eukaryotes (average 584 WD40 motifs and 193 WD40 motif-bearing proteins per genome). The number of WD40 motif-bearing proteins was found to correlate with genome size and gene number. Most WD40 motif-bearing proteins of Planctomycetes species belonged to the categories of 'ribosome assembly protein 4' and 'eukaryotic-like serine/threonine protein kinase.' Collinearity analysis of amino acid compositions of Planctomycetes and eukaryotic WD40 motifs revealed that the sequences of the four anti-parallel β-sheets of WD40 motifs were conserved. However, a number of Planctomycetes WD40 motifs had increased size of the interval region of β-sheets D and A. Taken together, results of this study suggest a positive correlation between the number of WD40 motif-bearing proteins and the evolution of Planctomycetes species toward a complex intracellular structure similar to that of eukaryotes.


Dose-Related Hemodynamic Effects of Enalaprilat in Patients with Ischemic Heart Disease.
To investigate the dose-related hemodynamic effects of angiotensin-converting enzyme inhibition (ACEI) in patients with angina pectoris (AP), five patients underwent right and left heart catheterization, and left ventriculogram before and 10 min after administration of 1.25 mg enalaprilat intravenously (Group 1). The results were compared with those of five patients with similar characteristics who received 2.5 enalaprilat (Group 2). There were no baseline differences between groups. After enalaprilat administration, there was significant differences between Groups 1 and 2, as follows: systolic blood pressure was 138 plus minus 16 versus 127 plus minus 14 mmHg (p = 0.05), left ventricular end-diastolic pressure was 15 plus minus 6 versus 7 plus minus 4 mmHg (p = 0.04), systemic vascular resistance was 1341 plus minus 290 versus 965 plus minus 271 dyne/sec/cm(minus sign5) (p = 0.05), pulmonary vascular resistance was 174 plus minus 39 versus 156 plus minus 15 dyne/sec/cm(minus sign5) (p = 0.05), end-diastolic volume was 87 plus minus 34 versus 107 plus minus 17 ml (p = 0.03), cardiac index was 3.0 versus 4.5 L min(minus sign1) m(minus sign2) (p = 0.01), left ventricle end-systolic wall stress was 31 plus minus 10 versus 22 plus minus 9 k dyne cm(minus sign5) (p = 0.01), ejection fraction was 69 plus minus 12 versus 81 plus minus 8% (p = 0.01), and wall motion index was 1.14 plus minus 0.1 versus 1.02 plus minus 0.1 (p = 0.001). These results indicate that enalaprilat has dose-related effects improving hemodynamics and ventricular function in patient with AP.


The Special Needs of Preterm Children - An Oral Health Perspective.
Preterm births are defined as those before 37 weeks of gestation. With advances in fertility medicine and neonatal medicine, the numbers of preterm children in the community have significantly increased. Developmental delays and complications among preterm children are well recognized. Much less consideration is given to the dental complications of preterm children. Manifestations include palatal deformations, enamel defects, tooth size variations and tooth shape deformities, malocclusions, and increased risks of early childhood caries and tooth wear. This article explores orodental risks and orodental needs of preterm children and suggests preventive and management strategies for optimizing the oral health of special needs children.


Long-Term Follow-up of Small Pulmonary Ground-Glass Nodules Stable for 3 Years: Implications of the Proper Follow-up Period and Risk Factors for Subsequent Growth.
How long persistent and stable ground-glass nodules (GGNs) should be followed is uncertain, although a minimum of 3 years is suggested. Here, we evaluated a group of GGNs that had remained stable for an initial period of 3 years with the aim of determining the proportion of GGNs showing subsequent growth after the initial 3 years and identifying the clinical and radiologic factors associated with subsequent growth. We retrospectively analyzed patients who underwent further computed tomography (CT) after the initial 3-year follow-up period showing a persistent and stable GGN (at least 5 years of follow-up from the initial CT). Between May 2003 and June 2015, 453 GGNs (438 pure GGNs and 15 part-solid GGNs) were found in 218 patients. Of the 218 patients, 14 had 15 GGNs showing subsequent growth after the initial 3 years during the median follow-up period of 6.4 years. For the person-based analysis, the frequency of subsequent growth of GGNs that had been stable during the initial 3 years was 6.7% (14 of 218). For the nodule-based analysis, the frequency was 3.3% (15 of 453). In a multivariate analysis, age 65 years or older (OR = 5.51, p = 0.012), history of lung cancer (OR = 6.44, p = 0.006), initial size 8 mm or larger (OR = 5.74, p = 0.008), presence of a solid component (OR = 16.58, p = 0.009), and air bronchogram (OR = 5.83, p = 0.015) were independent risk factors for subsequent GGN growth. For the individuals with GGNs having the aforementioned risk factors, the longer follow-up period is required to confirm subsequent GGN growth.


Occupational screening for sleep disorders in 12-h shift nurses using the Berlin Questionnaire.
The Berlin Questionnaire has been validated as a screening tool for sleep apnea in clinical samples, but no occupational studies have reported screening validity parameters for this instrument. The objectives of this pilot study were to describe the prevalence of sleep-disordered breathing symptoms in registered nurses and examine the validity of the Berlin Questionnaire to screen for sleep apnea in this chronically partially sleep-deprived group. Validity parameters for the Berlin Questionnaire are tabulated for published studies to 2012. Twenty-one female nurses working full time 12-h shifts underwent overnight, in-laboratory polysomnography to identify sleep disorders and completed a Berlin Questionnaire. By polysomnogram, the prevalence of sleep-disordered breathing [Respiratory Disturbance Index (RDI) ≥ 5] was 43 %, although by the Berlin Questionnaire only 24 % were deemed high risk. The sensitivity of the Berlin Questionnaire to detect high-risk subjects (RDI > 5) was 0.33, with a specificity of 0.83, a positive predictive value of 0.60, and negative predictive value of 0.63. Berlin criterion 3 (obesity or hypertension) performed the best for predicting sleep apnea in 12-h shift nurses. Although the Berlin Questionnaire produced valuable data about symptoms of sleep apnea in this population, it had a high proportion of false negatives. To improve its sensitivity for screening health care workers for sleep apnea, it must better capture symptoms specific to this population. Increasing the weighting of Berlin criterion 3 items should be considered to improve its psychometric properties.


The effects of energy and carbohydrate restriction in patients with chronic diabetes mellitus.
Thirty-five freshly presenting, diabetic patients received 5 hour, 100 g oral glucose tolerance tests when first seen and after a period of carbohydrate and energy restriction. After treatment, the significant improvement in glucose tolerance was accompanied by increased insulin secretion and lower concentrations of blood ketone bodies, lactate, glycerol, FFA, triglycerides, cholesterol and pre-beta lipoprotein. There were no significant changes in serum growth hormone or blood pyruvate concentrations. Improvement in glucose tolerance was greater in patients who were obese (greater than 115% of desirable body weight for height) on presentation and was related to the improvement in insulin secretion and the diminished lipolysis. An hypothesis to explain the changes in insulin secretion is prosposed. Eleven out of the 35 patients showed sufficient improvement in glucose tolerance to require no treatment other than diet.


[Expression of CXC chemokine ligand 16 and chemokine receptor 6 in periodontal tissues of patients with moderately severe periodontitis].
To investigate the expression and localization of CXCLl6 and its receptor CXCR6 in gingival tissues of patients with chronic periodontitis and analyze the roles in the occurrence and development of periodontitis. Gingival tissues with moderately severe periodontitis were collected from 39 patients treated in People's Hospital of Liaoning Province. Forty persons with healthy gingiva were used as control. The age of the enrolled subjects ranged from 35 to 74 years old. Immunohistochemical technique was applied to detect the expression and localization of CXCLl6 and CXCR6 protein gingival tissues of all subjects. The data was analyzed with SPSS 15.0 software package. Compared with normal group, the expression of CXCLl6 and CXCR6 were significantly up-regulated in patients with periodontitis (P<0.05). There was a positive relationship between the protein expression of CXCLl6 and CXCR6 in gingival tissues of patients with periodontitis (r=0.580, P<0.05). CXCLl6 and CXCR6 may be involved in the development of chronic periodontitis. The expression of CXCLl6 and CXCR6 may work coordinately to promote the development of chronic periodontitis.


Comparative study on three surgical techniques for intra-articular calcaneal fractures: open reduction with internal fixation using a plate, external fixation and minimally invasive surgery.
To evaluate, compare and identify the surgical technique with best results for treating intra-articular calcaneal fractures, taking into account postoperative outcomes, complications and scoring in the Aofas questionnaire. This was a retrospective study on 54 patients with fractures of the calcaneus who underwent surgery between 2002 and 2012 by means of the following techniques: (1) open reduction with extended L-shaped lateral incision and fixation with double-H plate of 3.5 mm; (2) open reduction with minimal incision lateral approach and percutaneous fixation with wires and screws; and (3) open reduction with minimal incision lateral approach and fixation with adjustable monoplanar external fixator. Patients treated using a lateral approach, with fixation using a plate had a mean Aofas score of 76 points; those treated through a minimal incision lateral approach with screw and wire fixation had a mean score of 71 points; and those treated through a minimal incision lateral approach with an external fixator had a mean score of 75 points. The three surgical techniques were shown to be effective for treating intra-articular calcaneal fractures, without any evidence that any of the techniques being superior. Intra-articular calcaneal fractures are complex and their treatment should be individualized based on patient characteristics, type of fracture and the surgeon's experience with the surgical technique chosen.


Treatment of symptomatic osteoporosis with fluoride.
Fluoride has been used for > 30 years in the management of osteoporosis. It is one of the few agents that has marked anabolic effects on the skeleton. Indeed, treatment results in continued increments in cancellous bone volume so that cancellous bone volume can be restored to normal in patients with advanced osteoporosis. Despite its long history, both the efficacy and risks of fluoride regimens continue to be the subject of controversy. One of its problems is its age, and much of the early work undertaken utilized methodology and proofs that are today unacceptable. A further problem is that sodium fluoride as a treatment is cheap and not patented (although some formulations are), so that it has been difficult for investigators or industry to make the investments required to provide a modern program of evaluation.


Tele-accelerometry as a novel technique for assessing functional status in patients with heart failure: feasibility, reliability and patient safety.
The six-minute walk test (6MWT) is an established measure of functional exercise capacity associated with clinical prognosis in Chronic Heart Failure (CHF). The aim of this study is to evaluate the employment of tele-accelerometry for the remote assessment of 6MWT in CHF. 155 patients were subjected to tele-accelerometry in form of monthly 6MWTs in their home surroundings. Accelerometer output included steps/6 min and walking speed. Data was directly transferred via mobile network to the Telemedicine Centre subsequent to test completion. 6MWT distance was measured by hand wheel and steps were counted with a digital hand-counter at baseline (Test 1) and at 12 months follow-up (Test 2). Accelerometer accuracy was within the 99th percentile. There was a significant correlation between step count, walking speed and measured 6MWT distance (Test 1: steps: r=0.80, P<0.001; Test 2: steps: r=0.90, P<0.001 and Walking Speed Test 1: r=0.80, P<0.001; Walking Speed Test 2: r=0.86, P<0.001). The reproducibility of tele-accelerometry was within 95% margin for all performance parameters, which showed stronger associations to quality of life questionnaire (Short Form - 36) Physical Component Score (PCS) than New York Heart Association (NYHA) functional class. Tele-accelerometry is feasible in patients with CHF and output parameters are indicative of exercise capacity. The benefit of this approach lies in its simplicity under every day circumstances by enabling routine performance testing to assess patients' functional status.


Adverse outcomes of endoscopic retrograde cholangiopancreatography: avoidance and management.
Adverse outcomes of endoscopic retrograde cholangiopancreatography (ERCP) include technical failures and complications. Complications cause significant morbidity to patients and anxiety to endoscopists. The key to preventing complications is to understand which patients and procedures are at highest risk so that appropriate decisions can be made as to whether ERCP should be performed at all, and if so, how. Patients who need ERCP the least are often the ones most likely to develop complications. For marginal indications, ERCP should be avoided. Success rates are higher and complication rates lower for endocopists performing large numbers by endoscopists with adequate experience.


Research on the mechanism of information infrastructure affecting industrial structure upgrading.
The construction of information infrastructure as well as the transformation and upgrading of the industrial structure are among the major challenges for the Chinese economy. Therefore, it is of great significance to explore how information infrastructure affects the upgrading of industrial structure. Based on the panel data of 31 provinces in China from 2013 to 2020, mediating effect model and non-parametric percentile bootstrap method are used to carry out empirical research, by creating an information infrastructure construction level and industrial structure upgrading indicators. The results show that, in addition to the direct effect of information infrastructure on industrial structure upgrading, information infrastructure can also work indirectly through three paths: (1) information infrastructure acts on industrial structure upgrading by enhancing urbanization level; (2) information infrastructure affects industrial structure upgrading by boosting technological innovation; (3) information infrastructure first enhances urbanization level, then acts on technological innovation, and finally promotes industrial structure upgrading. In addition, the intermediary effect of technological innovation is stronger than urbanization. In general, this study acknowledges that urbanization and technological innovation are partial mediators in the process of information infrastructure affecting industrial structure upgrading, notwithstanding other potential impact pathways being studied further.


Vascular endothelial growth factor-C: a growth factor for lymphatic and blood vascular endothelial cells.
The endothelial cells lining all vessels of the circulatory system have been recognized as key players in a variety of physiological and pathological settings. They act as regulators of vascular tone via the inducible nitric oxide system and in angiogenesis, the formation of blood vessels de novo. Aberrant regulation of endothelial cells contributes to tumor formation, atherosclerosis, and diseases such as psoriasis and rheumatoid arthritis. Among the most recently discovered growth factors for endothelial cells are newly isolated members of the platelet-derived growth factor/vascular endothelial growth factor (VEGF) family, VEGF-B, VEGF-C, and VEGF-D. VEGF-C is the ligand for the receptor tyrosine kinase VEGFR-3 (also known as Flt4), which is expressed predominantly in lymphatic endothelium of adult tissues, but a proteolytically processed form of VEGF-C can also activate VEGFR-2 of blood vessels. The lymphatic vessels have been known since the 17th century, but their specific roles in health and disease are still poorly understood. With the discovery of VEGF-C and its cognate receptor VEGFR-3, the regulation and functions of this important component of the circulatory system can be investigated.


Significance and prognostication of mediastinal lymph node enlargement on chest computed tomography among adult Indigenous Australians.
There is a lack of data on chest computed tomography (CT) findings on mediastinal lymph node enlargement (MLE), including normal size threshold of less than 10 or 15 mm for MLE among Indigenous Australians. In this study, we assessed the significance and the applicability of the current guidelines for the threshold for abnormal MLE among adult Indigenous Australians. Patients who underwent chest CT between 2012 and 2020 among those referred to undergo lung function test (spirometry) were assessed for the presence of MLE which were classified as Group A (no measurable nodes), Group B (<10 mm), Group C (≥10 to 14.99 mm) and Group D (≥15 mm). Of the total 67 patients identified to have MLE, 49 patients had at least two CT scans available for assessment over a median follow-up period of 101.3 weeks (IQR: 62.4, 235.6) and were included in the analysis. Evidence of chronic lung disease was common, with a significant proportion demonstrating either COPD or bronchiectasis and a high proportion with smoking history (93%). During the first CT scan, 34/49 (69%) had >10 mm nodes, of which 12/34 (35%) reduced in size, 22/34 (65%) remained stable, and 3/34 (9%) had malignancy on follow-up. Despite most patients demonstrating the presence of significant MLE with varying size and in most >10 mm, the majority remain stable or benign in nature and only a minor proportion showed evidence of lung malignancy. Further prospective studies are needed in the characterisation of MLE among Indigenous patients.


Involvement of nitric oxide and potassium channels in the bradykinin-induced vasodilatation in the rat kidney perfused ex situ.
The role of nitric oxide (NO), K(+) channels, and arachidonic acid metabolism, via cytochrome P450 and cyclooxygenase pathways, in the renal vasodilatory effect of bradykinin was examined in the isolated rat kidney perfused ex situ with a blood-free solution. Bradykinin (BK, 0.25-1.0 microM) induced a dose-dependent reduction of 10-35% in the relative renal vascular resistance (rRVR) of isolated kidneys preconstricted with phenylephrine (PHE, 0.17-0.35 microM). The vasodilating effect of 0.5 microM bradykinin was significantly inhibited by the nitric oxide synthase inhibitors, N(G)-nitro-L-arginine (95% inhibition) and N(G)-nitro-L-arginine methyl ester (45-75% inhibition). Clotrimazole, an inhibitor of cytochrome P450 pathway but not indomethacin, a cyclooxygenase inhibitor, reduced the renal vasodilator response to bradykinin by 84%. The nonspecific K(+) channel inhibitor, tetraethylammonium ion (TEA) and the selective inhibitor of Ca(2+)-activated K(+) channels, charybdotoxin (ChTX) greatly attenuated the vasodilator response to bradykinin by approximately 84% and 79%, respectively. These two K(+) channel inhibitors showed similar effects on vasodilatation induced by S-nitroso-acetyl-D,L-penicillamine (1 microM), a nitric oxide donor. The results suggest that bradykinin releases nitric oxide which, by opening potassium channels specifically the Ca(+)-dependent type, mediates the renal vasodilator response to bradykinin in the isolated kidney perfused ex situ.


Gut microbial metabolite p-cresol alters biotransformation of bisphenol A: Enzyme competition or gene induction?
Recent studies on pharmaceuticals have revealed the direct and indirect mechanisms that link human gut microbiome to xenobiotic biotransformation. Though environmental contaminants compose a vital portion of xenobiotics and share overlapping biotransformation pathways with gut microbial metabolites, the possible interplay between gut microbiome and biotransformation of environmental contaminants remains obscure. This study utilized bisphenol A (BPA) and p-cresol as model compounds to explore whether gut microbial metabolites could affect environmental phenol metabolism on both in vitro and in vivo models. We have observed some distinct biotransformation behavior, where in vivo mouse examination using 171 & 1972 μg/kg bw p-cresol injection exhibited enhancing effect on BPA metabolism, but p-cresol was found as a strong inhibitor from 10/5 μM in a non-competitive pattern for BPA biotransformation in in vitro models of liver S9 fractions and HepG2 cell line, respectively. A further investigation revealed that the expression of biotransformation enzyme genes including Ugt1a1, Ugt2b1, or Sult1a1 of p-cresol treated mice were dynamically induced. In silico docking approach was also utilized to explore the non-competitive inhibition mechanism by estimating the binding affinity of key enzyme SULT 1A1. Overall, our results provided a novel insight into the biotransformation interaction between gut microbiome and environmental contaminants.


Quantitative determination of 5-hydroxyindole-3-acetic acid in body fluids by high-performance liquid chromatography.
A method for the quantitative determination of 5-hydroxyindole-3-acetic acid in urine and cerebrospinal fluid is described. The method is based on reversed phase high-performance liquid chromatography and fluorometric detection. The experimental error was below 5% in urine and 8% in cerebrospinal fluid. Urine and cerebrospinal fluid samples were analysed both by the described method and by a mass fragmentographic method. The correlation coefficients for the two methods were 0.986 in the urine samples and 0.965 in the cerebrospinal fluid samples. Analysis of 100 samples can be achieved within 3 days. The described method offers a possibility to determine 5-hydroxyindole-3-acetic acid in urine and cerebrospinal fluid with the same precision and capacity as do mass fragmentography but with the use of a less sophisticated equipment.


Pigment Epithelium-Derived Factor and Sex Hormone-Responsive Cancers.
Oestrogens and androgens play important roles in normal and cancerous tissue and have been shown to negatively regulate pigment epithelium-derived factor (PEDF) expression in sex hormone-responsive tumours. PEDF suppresses tumour growth and its downregulation by oestrogen is implicated in tumorigenesis, metastasis, and progression. PEDF expression is reduced in cancerous tissue of the prostate, breast, ovary, and endometrium compared to their normal tissue counterparts, with a link between PEDF downregulation and sex hormone signalling observed in pre-clinical studies. PEDF reduces growth and metastasis of tumour cells by promoting apoptosis, inhibiting angiogenesis, increasing adhesion, and reducing migration. PEDF may also prevent treatment resistance in some cancers by downregulating oestrogen receptor signalling. By interacting with components of the tumour microenvironment, PEDF counteracts the proliferative and immunosuppressive effects of oestrogens, to ultimately reduce tumorigenesis and metastasis. In this review, we focus on sex hormone regulation of PEDF's anti-tumour action in sex hormone-responsive tumours.


Genetic linkage between the loci for myotonic dystrophy and peptidase D.
In a linkage study between myotonic dystrophy and peptidase D it is evident from the lod score values that with high probability theta lies between 0 and 0.1. The data thus support a previous hint of linkage between peptidase D and the Lutheran and secretor loci, which were already known to be linked to myotonic dystrophy.


Reduced expression and defective modulation of TNF receptor/ligand family molecules on proB-ALL blasts.
There is a subgroup of pediatric patients with an immature immunophenotype of proB-ALL that still poses a therapeutic challenge, even if the overall prognosis in B cell precursor acute lymphoblasic leukemia (BCP-ALL) is very good. Due to impaired treatment response these patients are prone to suffer relapse and are thus by definition stratified into the clinically defined high risk group receiving intensified chemotherapy. Besides response to chemotherapy long term prognosis is also influenced by immunological control mechanisms. Thus, high expression of the TNF receptor CD40 has been shown to prevent particularly late relapse in BCP-ALL suggesting a pivotal role of this regulatory molecule for maintenance of the remission status. We therefore determined the baseline expression and CD40-mediated modulation of TNF receptor and costimulatory molecules in 5 patients with proB-ALL, 8 with preB-ALL and 22 with c-ALL performing FACS analysis. We particularly compared the TNF receptor status on proB-ALL blasts to the expression on more mature preB- and c-ALL blasts. Here, we demonstrate for the first time a significantly lower baseline expression and CD40-induced modulation capacity of TNF receptor and costimulatory molecules in pediatric proB-ALL compared to more mature precursor B-ALL blasts. The lower expression and defective capacity of proB-ALL blasts to respond to CD40 ligand stimulation might resemble the immature feature of these blasts and besides increased chemoresistance contribute to the impaired prognosis of these patients due to escape from apoptosis and immunological control mechanisms.


[Cervical cerclage in Reunion island: Evaluation of physicians' practice patterns].
Cervical incompetency is one of the direct causes of neonatal morbidity and mortality; a unique and efficient treatment of which is cervical cerclage. The objective of this study was the evaluation of physicians' practice patterns concerning cerclage in Reunion Island, in order to reinforce the management and information of patients at risk. The indications and complications of cerclages effectuated in 2010 and 2011 were compared to the literature. In this retrospective study, all the medical records of cerclage realized in Reunion Island during two years were collected and analyzed, specifically data concerning patients' cerclage, the complications, and the outcome of the pregnancy. We listed 200 cerclages, which were predominantly prophylactic cerclages (75.5%) and represented 0.71% of all births. A total of 71% of the indications of cerclage in Reunion Island did not take into account the recommendations of the literature. Analysis revealed the frequent use of prophylactic cerclage and subsequently reflected the insufficient use of therapeutic cerclage. In those cases, the rate of premature delivery was indeed lower (P=0.003), as well as the rate of chorioamniotitis (P=0.003). Cerclage is an efficient treatment to extend the length of the pregnancy. Nevertheless, it is important to comply with the recommendations given by the literature, by spotting the patients at risk of premature delivery, and recommend cerclage only in case of real cervical incompetency, for the sake of improving their management and reducing the rate of complications.


Tensile strength of a new suture for fixation of tendon grafts when using a weave technique.
To evaluate a new corner stitch construct for tendon graft or tendon transfer fixation and compare the tensile strength with a conventional central cross-suture design in human cadaver tendons. Flexor digitorum profundus tendons of the index, middle, and ring fingers (48 total) were used as recipients and palmaris longus, extensor indicis proprius, and extensor digitorum communis tendons of the index finger (48 total) were used as grafts from 16 fresh-frozen human cadaver hands. We compared the cross-stitch technique with a new corner stitch technique in tendon repairs made with 1, 2, or 3 weaves (8 per group). Tendons were sutured at each weave with either 2 full-thickness cross-stitches or 4 partial-thickness corner stitches of 4-0 nylon. Mattress sutures also were placed through the free tendon end for each repair type. The tensile strength of the tendon-graft composite was measured with a materials testing machine. The tensile strength of the repairs increased significantly with the number of weaves. When 2 or 3 weaves were used with the corner stitch or when 3 weaves were used with the cross-stitch, the repairs were significantly stronger. Although no significant difference in strength to failure was noted when comparing cross and corner stitches with equivalent numbers of weaves, qualitatively there was a difference in mode of failure with the 3-weave corner stitches failing primarily by intrasubstance tendon failure and the 3-weave cross-stitch repairs failing by tendon pullout. The corner stitch is as strong as conventional cross-stitch repairs and its superficial placement may be more favorable to tendon blood supply. This repair may be advantageous for clinical applications.


Intrarectal administration of mCRAMP-encoding plasmid reverses exacerbated colitis in Cnlp(-/-) mice.
Cathelicidin is a pleiotropic host defense peptide secreted by epithelial and immune cells. Whether endogenous cathelicidin is protective against ulcerative colitis, however, is unclear. Here we sought to delineate the role of endogenous murine cathelicidin (mCRAMP) and the therapeutic efficacy of intrarectal administration of mCRAMP-encoding plasmid in ulcerative colitis using dextran sulfate sodium (DSS)-challenged cathelicidin-knockout (Cnlp(-/-)) mice as a model. Cnlp(-/-) mice had more severe symptoms and mucosal disruption than the wild-type mice in response to DSS challenge. The tissue levels of interleukin-1β and tumor necrosis factor-α, myeloperoxidase activity and the number of apoptotic cells were increased in the colon of DSS-challenged Cnlp(-/-) mice. Moreover, mucus secretion and mucin gene expression were impaired in Cnlp(-/-) mice. All these abnormalities were reversed by the intrarectal administration of mCRAMP or mCRAMP-encoding plasmid. Taken together, endogenous cathelicidin may protect against ulcerative colitis through modulation of inflammation and mucus secretion.


Volar locking plate fixation of distal radius fractures: Isokinetic assessment of the influence of an ulnar styloid process fracture on pronation-supination strength.
Although distal radius fractures (DRF) are frequent, the management of associated ulnar styloid process (USP) lesions is still controversial. According to recent studies, a fracture of the USP does not appear to affect functional outcomes after treatment of a DRF with plate fixation. We sought to compare the impact of a USP fracture on pronation and supination strength in isometric and isokinetic tests. We included patients with a DRF who underwent volar locking plate fixation. We divided our population into three groups: one group consisted of patients who had a fracture of the USP base, one group composed of patients without USP fracture or with a distal ulnar fracture only, and a control group composed of subjects with normal wrists. Inclusion criteria included an age of 18 to 50 years and a minimum follow-up of 10 months post-surgery. The main exclusion criteria were complex intraarticular fractures and postoperative complications. The assessment was based on clinical examination and recording of forearm rotation strength during pronation and supination in isokinetic and isometric tests. The ratio between the operated and contralateral sides was compared for each patient. Thirty-six participants were included (mean age 31.1±4.5 years). The mean postoperative follow-up was 17.9±6.9 months. Participants with a USP fracture differed from other participants in their peak torque on the isokinetic test at 45°/s for pronation and supination, but not on isokinetic tests at 120°/s or in isometric tests. Isokinetic tests revealed a decrease in pronation-supination strength during sustained effort for patients with associated basal USP fractures. These findings may have clinical implications for management of the USP but need to be specified with further study. LEVEL OF EVIDENCE: prognostic study level III.


CRISPR: A new paradigm of theranostics.
Infectious and hereditary diseases are the primary cause of human mortality globally. Applications of conventional techniques require significant improvement in sensitivity and specificity in therapeutics. However, clustered regularly interspaced short palindromic repeats (CRISPRs) is an innovative genome editing technology which has provided a significant therapeutic tool exhibiting high sensitivity, fast and precise investigation of distinct pathogens in an epidemic. CRISPR technology has also facilitated the understanding of the biology and therapeutic mechanism of cancer and several other hereditary diseases. Researchers have used the CRISPR technology as a theranostic approach for a wide range of diseases causing pathogens including distinct bacteria, viruses, fungi and parasites and genetic mutations as well. In this review article, besides various therapeutic applications of infectious and hereditary diseases we have also explained the structure and mechanism of CRISPR tools and role of CRISPR integrated biosensing technology in provoking diagnostic applications.


The Ito "Flow-Through" Centrifuge. A new device for long-term (24 hours) plasmapheresis without platelet deterioration.
An efficacious procedure for continuous plasmapheresis in long-term extracorporeal circulation has not yet been devleoped. This technique could be important in plasma exchange or cross-exchange and in artificial organ support if it could be accomplished safely using heparin as the anticoagulant. When sheep were connected to the Celltrifuge for 23 hours and administered 150 units/kg per hour heparin anticoagulant, we found gross platelet clumping in separated plasma and a 60 per cent fall in platelet count. When 120,000 units of heparin were directly injected into the centrifuge bowl at the onset of the centrifugation followed by hourly heparin administration at 500 units/kg, there was no platelet clumping but the platelet count fell to 52 per cent. Under identical conditions the Ito Flow-Through Centrifuge, a new continuous flow centrifuge with no rotating seals, was used. We found no platelet aggregates and no drop in platelet count, and there was a 100 per cent recovery of platelets in the separated plasma. Platelets sampled from arterial blood and separated plasma were found to have normal function in vitro, and responded 100 per cent to both collagen and ADP. These results indicate that the Ito Flow-Through Centrifuge can be used in plasmapheresis with heparin anticoagulation with no donor platelet loss and normal function in separated plasma.


Caspase-like proteases and the phytohormone cytokinin as determinants of S-RNAse-based self-incompatibility-induced PCD in Petunia hybrida L.
S-RNAse-based self-incompatibility (SI) in petunia (Petunia hybrida L.) is a self-/non-self-recognition system underlying the pistil rejection of self-pollen. Using different methods, including a TUNEL assay, we have recently shown that programmed cell death (PCD) is a factor of the SI in petunia. Here, we show that the growth of self-incompatible pollen tubes in the style tissues during 4 h after pollination is accompanied by five-sixfold increase in a caspase-like protease (CLP) activity. Exogenous cytokinin (CK) inhibits the pollen tube growth and stimulates the CLP activity in compatible pollen tubes. The actin depolymerization with latrunculin B induces a sharp drop in the CLP activity in self-incompatible pollen tubes and its increase in compatible pollen tubes. Altogether, our results suggest that a CLP is involved in the SI-induced PCD and that CK is a putative activator of the CLP. We assume that CK provokes acidification of the cytosol and thus promotes the activation of a CLP. Thus, our results suggest that CK and CLP are involved in the S-RNAse-based SI-induced PCD in petunia. Potential relations between these components in PCD signaling are discussed.


Attainment of personal goals in the first year of intrathecal baclofen treatment in dyskinetic cerebral palsy: a prospective cohort study.
To assess attainment of individual treatment goals one year after intrathecal baclofen (ITB) pump implantation in individuals with dyskinetic cerebral palsy (CP). A multi-center prospective cohort study was conducted including 34 non-walking individuals with severe dyskinetic CP, classified as Gross Motor Function Classification System (GMFCS) IV/V, aged 4-24 years, 12 months after pump implantation. The main outcome measure was Goal Attainment Scaling (GAS). Predictors of GAS results were analyzed. Complications were registered systematically. Seventy-one percent of individuals with dyskinetic CP fully achieved one or more treatment goals. One or more treatment goals were partially achieved in 97% of individuals. Two factors were found to be associated with attainment of goals: Dyskinesia Impairment Scale (DIS) score at baseline and the difference in pain score between baseline and follow-up. These two variables explain 30% of the variance in the outcome. Intrathecal baclofen is effective in achieving individual treatment goals in children and young adults with dyskinetic CP after nine to 12 months of ITB treatment. A positive outcome on treatment goals is, for a small part, associated with higher severity of dystonia at baseline and with improvement of pain during treatment. Dutch Trial Register, number NTR3642.Implications for rehabilitationIntrathecal baclofen treatment is effective in attainment of personal treatment goals, one year after pump implantation in patients with dyskinetic cerebral palsy.A positive outcome on treatment goals is, for a small part, related to higher severity of dystonia at the start and on improvement of pain during treatment.


Pharmacokinetic and pharmacodynamic properties of SOL1: a novel dual inhibitor of neutral endopeptidase and endothelin converting enzyme.
The pharmacological profile of the novel putative neutral endopeptidase (NEP) and endothelin converting enzyme (ECE) inhibitor SOL1 was examined. The enzyme inhibitory profile of SOL1 was established in vitro. The pharmacokinetic and pharmacodynamic profile was determined in rodents in vivo. In vitro, at neutral pH, 10 μM SOL1 inhibited NEP-1, NEP-2, and ECE-1 by 99%, 94% and 75%, respectively. The IC(50)s were 25, 25 and 3200 nmol/L, respectively. In anesthetized rats, SOL1 inhibited blood pressure (BP) responses to big-ET-1 and ET-1(1-31) with ED(50)s of 1.9 and 0.03 mg/kg, corresponding to plasma EC(50)s of 4.6 and 0.1 μmol/L, respectively. Pharmacokinetics of SOL1 were examined after single injections in mice and rats. In these species, the estimated clearance of SOL1 varied between 5 and 9 ml/kg.min and T(1/2) between 20 and 60 min. Steady state kinetics of SOL1 were examined after continuous s.c. infusions of SOL1 for 3 weeks at 50mg/kg.day in DOCA-salt hypertensive rats. This treatment lowered BP by 22 mmHg. Steady state concentrations of SOL1 in plasma were 3.9 μmol/L. In heart, lung, and kidney the concentrations of SOL1 were 0.4, 1.8, and 20.5 μmol/kg, respectively. About 63% of the daily dose was retrieved unaltered in the urine. These data indicate that SOL1 is primarily a NEP inhibitor in vitro as well as in vivo. Given the preferential renal accumulation and renal clearance of SOL1 additional ECE-1 inhibition in the kidney may have contributed to its chronic BP lowering effects in the DOCA-salt hypertensive rat model.


Lectins detect changes of the glycosylation status of plasma membrane constituents during late apoptosis.
Mechanisms governing the normal resolution processes of inflammation are poorly understood, yet their elucidation may lead to a greater understanding of the pathogenesis of chronic inflammation. The removal of apoptotic cell material and their potentially histotoxic contents is a prerequisite of resolution. Engulfment by macrophages is an important disposal route, and changes in the apoptotic cells that are associated with their recognition by macrophages are the subject of this report. Apoptosis and necrosis in primary cells and cell lines were induced by various stimuli. The binding profile of 23 different lectins for vital, apoptotic, and necrotic cells were analyzed by flow cytometry. We observed that lectins were able to attach to the cell surfaces of vital and dying cells. Some lectins exhibited membrane destructive properties and, consecutively, changed the morphology of the cells as detected by flow cytometry. Other lectins did not show differences in their binding to viable and apoptotic cells. Those lectins were, therefore, not used for analyses of surface changes. The lectins Griffonia simplificolia II (GSL II), Narcissus pseudonarcissus (NPn), and Ulex europaeus I (UEA I) showed no cytotoxic activity and bound preferentially to dying cells. Primary and secondary necrotic cells displayed an equal staining intensity, which was substantially higher than for apoptotic cells. The binding of GSL II, NPn, and UEA to dying cells increased in a time-dependent manner and was delayed to AxV positivity and the decrease in the mitochondrial membrane potential of apoptotic cells. The kinetic of the lectin staining correlated with the increase in subG1-DNA. GSL II, NPn, and UEA are specific for N-acetylglucosamine, mannose, and fucose, respectively. According to their binding specificity, we conclude that N-acetylglucosamine-, mannose-, and fucose-containing epitopes are increasingly exposed on cells undergoing apoptosis.


Region-specific alterations in tyrosine hydroxylase activity in rats exposed to lead.
Previous studies from our laboratory showed that subchronic exposure to low levels of Pb resulted in significant decrease in dopamine (DA) content, attenuation of stimulus-induced release of DA in the dopaminergic projection area of nucleus accumbens (NA), and alterations in tyrosine hydroxylase (TH) activity in rat whole brain homogenates. The present study reported here was conducted to assess the functional integrity of DA synthesis in different brain regions of rats subchronically (90-days) exposed to 50 ppm Pb by measuring the activity of the rate limiting enzyme, tyrosine hydroxylase, in seven brain regions. In Pb-exposed rats, TH activity was reduced in two of the seven brain regions investigated, i.e., nucleus accumbens (42% reduction) and frontal cortex (61% reduction) when compared to controls. In contrast, Pb exposure did not affect the TH activity in cerebellum, brainstem, hippocampus, hypothalamus and striatum. The changes in TH activity in nucleus accumbens (NA) and frontal cortex (FC) in Pb-exposed rats were further confirmed by Western blot analysis using TH polyclonal antibody. Collectively, these results indicate that low level subchronic Pb exposure may affect TH protein in these brain regions.


Experience and outcomes of micrografting for major paediatric burns.
The deficit of donor sites in major burns over 50% of the total body surface area has necessitated the application of methods besides traditional meshed autografting to achieve definitive skin cover. The Meek micrografting technique was introduced at this hospital in 2011, especially in the absence of a reliable source of deceased donor allograft skin. The purpose of this study was to evaluate this strategy with reference to its technical execution, efficacy and indications in the context of major paediatric burn surgery. A cohort study was performed of all paediatric patients with major burn who underwent Meek micrografting at a dedicated paediatric burn centre in a developing country over a five year period. Demographics, details of their burns, operative management and clinical course and outcomes were collected from patient records and operative notes and analysed. Thirty-five patients were managed using the micrografting technique during the study period. The mean patient age was 4.1 years (range 3 months-11 years) and their mean total body surface area (TBSA) burn was 49.7% (range 15-86%). Eleven patients sustained inhalation injuries and five developed a re-feeding syndrome on account of delayed referral. The mean abbreviated burn severity index (ABSI) was 8.5 (range 2-13). The hospital length of stay in the 27 survivors was a mean of 75.5 days, equating to 1.4 days per percentage burn. Eight patients died during the course of treatment, with a mean TBSA burn of 67.75% (range 38-86%). Graft take one month after surgery was documented to be more than 90% in 24 patients, of whom 3 subsequently died. Eleven patients had less than 90% graft take at this time, of whom 5 died. There is a considerable 'learning curve' associated with this technique. In order to achieve success one must ensure a completely viable, non-infected bed, obtained by tangential or fascial excision, followed by allografting as temporary coverage and to 'test the wound bed' for definitive coverage. Infection resulted in the majority of autograft loss in this series, and in addition to risk factors like burn size and inhalation injury, accounted for many of the deaths in this series. Meek micrografting offers high expansion ratios, thereby facilitating durable wound cover in the presence of limited donor sites. It is unlikely that a lethal dose, 50% (LD50) of almost 70% TBSA would have been possible in this context without the regular application of this technique. This study advocates for the widespread availability of Meek micrografting and deceased donor allograft skin in developing countries.


Radiative heat conductances between dielectric and metallic parallel plates with nanoscale gaps.
Recent experiments have demonstrated that radiative heat transfer between objects separated by nanometre-scale gaps considerably exceeds the predictions of far-field radiation theories. Exploiting this near-field enhancement is of great interest for emerging technologies such as near-field thermophotovoltaics and nano-lithography because of the expected increases in efficiency, power conversion or resolution in these applications. Past measurements, however, were performed using tip-plate or sphere-plate configurations and failed to realize the orders of magnitude increases in radiative heat currents predicted from near-field radiative heat transfer theory. Here, we report 100- to 1,000-fold enhancements (at room temperature) in the radiative conductance between parallel-planar surfaces at gap sizes below 100 nm, in agreement with the predictions of near-field theories. Our measurements were performed in vacuum gaps between prototypical materials (SiO2-SiO2, Au-Au, SiO2-Au and Au-Si) using two microdevices and a custom-built nanopositioning platform, which allows precise control over a broad range of gap sizes (from <100 nm to 10 μm). Our experimental set-up will enable systematic studies of a variety of near-field-based thermal phenomena, with important implications for thermophotovoltaic applications, that have been predicted but have defied experimental verification.


Tumor-associated macrophages in lung carcinoma: From mechanism to therapy.
Tumor-associated macrophages (TAMs), which could be classified into the classical (M1-like) and alternatively activated (M2-like) phenotype, were considered to be important tumor-promoting components in lung cancer microenvironment. Several studies reported that TAMs in lung tumor islet or stroma are usually correlated with poor prognosis. Further studies showed that TAMs could promote the initiation of tumor cells, inhibit antitumor immune responses, and stimulate tumor angiogenesis and subsequently tumor metastasis of lung carcinoma. Currently, TAMs have been considered as penitential targets of lung cancer. This review summarizes from the fundamental information of TAMs to the its role in metastasis and present evidence for TAMs as a potential target of cancer therapy.


Cytokine related therapies for autoimmune disease.
In short, manipulation of cytokine pathways shows promise as a mean to tilt the balance of immunity toward tolerance. Effective and regulatory T cells vary in their response to a variety of cytokines. In particular, the ability of certain cytokines, for example, IL-2, to provide vital survival signals to regulatory cells and to trigger death of effector T cells or impede IL-15 driven expansion of memory cells has spurred several trials. The ability of IFNgamma, IL-4, TNFalpha, and lymphotoxin to exert selective effects upon crucial lymphocyte subset populations in vivo may also enable translation into potent therapies.


Network-wide thermodynamic constraints shape NAD(P)H cofactor specificity of biochemical reactions.
The ubiquitous coexistence of the redox cofactors NADH and NADPH is widely considered to facilitate an efficient operation of cellular redox metabolism. However, it remains unclear what shapes the NAD(P)H specificity of specific redox reactions. Here, we present a computational framework to analyze the effect of redox cofactor swaps on the maximal thermodynamic potential of a metabolic network and use it to investigate key aspects of redox cofactor redundancy in Escherichia coli. As one major result, our analysis suggests that evolved NAD(P)H specificities are largely shaped by metabolic network structure and associated thermodynamic constraints enabling thermodynamic driving forces that are close or even identical to the theoretical optimum and significantly higher compared to random specificities. Furthermore, while redundancy of NAD(P)H is clearly beneficial for thermodynamic driving forces, a third redox cofactor would require a low standard redox potential to be advantageous. Our approach also predicts trends of redox-cofactor concentration ratios and could facilitate the design of optimal redox cofactor specificities.


The effect of a knee ankle foot orthosis incorporating an active knee mechanism on gait of a person with poliomyelitis.
The aim of this case study was to identify the effect of a powered stance control knee ankle foot orthosis on the kinematics and temporospatial parameters of walking by a person with poliomyelitis when compared to a knee ankle foot orthosis. A knee ankle foot orthosis was initially manufactured by incorporating drop lock knee joints and custom molded ankle foot orthoses and fitted to a person with poliomyelitis. The orthosis was then adapted by adding electrically activated powered knee joints to provide knee extension torque during stance and also flexion torque in swing phase. Lower limb kinematic and kinetic data plus data for temporospatial parameters were acquired from three test walks using each orthosis. Walking speed, step length, and vertical and horizontal displacement of the pelvis decreased when walking with the powered stance control knee ankle foot orthosis compared to the knee ankle foot orthosis. When using the powered stance control knee ankle foot orthosis, the knee flexion achieved during swing and also the overall pattern of walking more closely matched that of normal human walking. The reduced walking speed may have caused the smaller compensatory motions detected when the powered stance control knee ankle foot orthosis was used. The new powered SCKAFO facilitated controlled knee flexion and extension during ambulation for a volunteer poliomyelitis person.


Multiple genetic lineages challenge the monospecific status of the West African endemic frog family Odontobatrachidae.
Correct species identification is crucial in different fields of biology, and in conservation. The endemic West African frog family Odontobatrachidae currently contains a single described species, Odontobatrachus natator. From western Guinea to western Côte d'Ivoire it inhabits forests around waterfalls or cascades. Based on more than 130 specimens from 78 localities, covering the entire distribution, we investigated the molecular diversity of these frogs. Our analyses included mitochondrial and nuclear genes, with a concatenated alignment of 3527 base pairs. We detected high level of genetic differentiation with five distinct lineages or operational taxonomic units (OTUs). These OTUs were also identified by two different species delimitation approaches, Generalized Mixed Yule Coalescent (GMYC) and cluster algorithm. All OTUs occur in parapatry in the Upper Guinean forests. One OTU, assigned to the "true" Odontobatrachus natator, covers the largest distribution, ranging from the border region of western Sierra Leone-Guinea to eastern Liberia. Two OTUs are restricted to western Guinea (Fouta Djallon and foothills), while two others occur in eastern Guinea and the border region of Guinea-Liberia-Côte d'Ivoire. The OTU representing O. natator consists of two divergent subclades: one restricted to the Freetown Peninsula (Sierra Leone) and the other covering all populations further inland. Environmental niche models indicated that the restricted Freetown Peninsula population is separated by unsuitable habitat from remaining populations. Geographic isolation of OTUs and molecular differences comparable to species level differentiation in other frog families indicate that O. natator contains cryptic species diversity. Respective distribution patterns most probably resulted from repeated changes of forest cover (contraction and expansion) over evolutionary timescales. The survival within forest refugia that have persisted through multiple drier periods and subsequent dispersal during wetter times may best explain the observed geographic distributions of OTUs. According to the IUCN Red List range criteria each OTU should be classified as "Endangered." If the Freetown Peninsula "natator" population is recognized as a distinct species it would warrant recognition as "Critically Endangered." The identification of cryptic lineages highlights the urgent need to protect these frogs, all of which are endemic to small areas within the Upper Guinean biodiversity hotspot.


Evaluation of Temporal Trends in Racial and Ethnic Disparities in Sleep Duration Among US Adults, 2004-2018.
Historically marginalized racial and ethnic groups are generally more likely to experience sleep deficiencies. It is unclear how these sleep duration disparities have changed during recent years. To evaluate 15-year trends in racial and ethnic differences in self-reported sleep duration among adults in the US. This serial cross-sectional study used US population-based National Health Interview Survey data collected from 2004 to 2018. A total of 429 195 noninstitutionalized adults were included in the analysis, which was performed from July 26, 2021, to February 10, 2022. Self-reported race, ethnicity, household income, and sex. Temporal trends and racial and ethnic differences in short (<7 hours in 24 hours) and long (>9 hours in 24 hours) sleep duration and racial and ethnic differences in the association between sleep duration and age. The study sample consisted of 429 195 individuals (median [IQR] age, 46 [31-60] years; 51.7% women), of whom 5.1% identified as Asian, 11.8% identified as Black, 14.7% identified as Hispanic or Latino, and 68.5% identified as White. In 2004, the adjusted estimated prevalence of short and long sleep duration were 31.4% and 2.5%, respectively, among Asian individuals; 35.3% and 6.4%, respectively, among Black individuals; 27.0% and 4.6%, respectively, among Hispanic or Latino individuals; and 27.8% and 3.5%, respectively, among White individuals. During the study period, there was a significant increase in short sleep prevalence among Black (6.39 [95% CI, 3.32-9.46] percentage points), Hispanic or Latino (6.61 [95% CI, 4.03-9.20] percentage points), and White (3.22 [95% CI, 2.06-4.38] percentage points) individuals (P < .001 for each), whereas prevalence of long sleep changed significantly only among Hispanic or Latino individuals (-1.42 [95% CI, -2.52 to -0.32] percentage points; P = .01). In 2018, compared with White individuals, short sleep prevalence among Black and Hispanic or Latino individuals was higher by 10.68 (95% CI, 8.12-13.24; P < .001) and 2.44 (95% CI, 0.23-4.65; P = .03) percentage points, respectively, and long sleep prevalence was higher only among Black individuals (1.44 [95% CI, 0.39-2.48] percentage points; P = .007). The short sleep disparities were greatest among women and among those with middle or high household income. In addition, across age groups, Black individuals had a higher short and long sleep duration prevalence compared with White individuals of the same age. The findings of this cross-sectional study suggest that from 2004 to 2018, the prevalence of short and long sleep duration was persistently higher among Black individuals in the US. The disparities in short sleep duration appear to be highest among women, individuals who had middle or high income, and young or middle-aged adults, which may be associated with health disparities.


Progesterone--specific binding sites in the kidney of the female baboon.
The uptake and retention of a radiolabeled synthetic progestin, ORG 2058, was studied in the urinary tract of the female baboon. Four estrogen-primed baboons were injected intravenously with 2.5 micrograms./kg. body weight of 3H-ORG 2058. One animal, which served as a control, received an additional injection of 2.5 mg./kg. body weight of unlabeled progesterone. One hour after the injections, the animals were killed and the kidneys, ureters and urinary bladder were removed and processed for autoradiography. Localization of progestin was observed in the nuclei of the convoluted and straight segments of the distal tubule, the ascending thick limb of the loop of Henle and both cortical and medullary collecting tubules. Connective tissue cells were also labeled in the medulla and cortex of the kidney. An absence of silver grains was noted in the renal corpuscle, all segments of the proximal tubule and the thin loop of Henle. Concentration of the tritiated steroid was not observed in either the ureter or bladder or in any portions of the urinary tract of the control animal. This study suggests that progesterone has a direct effect via a progesterone specific receptor on the various target cells that sequestered the 3H-ORG 2058.


The use of newborn foot length to identify low birth weight and preterm babies in Papua New Guinea: A diagnostic accuracy study.
Low birth weight (LBW, <2.50 kg) and preterm birth (PTB, <37 completed weeks of gestation) are important contributors to neonatal death. Newborn foot length has been reported to identify LBW and PTB babies. The objectives of this study were to determine the diagnostic accuracy of foot length to identify LBW and PTB and to compare foot length measurements of a researcher with those of trained volunteers in Papua New Guinea. Newborn babies were enrolled prospectively with written informed consent from their mothers, who were participating in a clinical trial in Madang Province. The reference standards were birth weight, measured by electronic scales and gestational age at birth, based on ultrasound scan and last menstrual period at the first antenatal visit. Newborn foot length was measured within 72 hours of birth with a firm plastic ruler. Optimal foot length cut-off values for LBW and PTB were derived from receiver operating characteristic curve analysis. Bland-Altman analysis was used to assess inter-observer agreement. From 12 October 2019 to 6 January 2021, we enrolled 342 newborns (80% of those eligible); 21.1% (72/342) were LBW and 7.3% (25/342) were PTB. The area under the curve for LBW was 87.0% (95% confidence intervals 82.8-90.2) and for PTB 85.6% (81.5-89.2). The optimal foot length cut-off was <7.7 cm for both LBW (sensitivity 84.7%, 74.7-91.2, specificity 69.6%, 63.9-74.8) and PTB (sensitivity 88.0% (70.0-95.8), specificity 61.8% (56.4-67.0). In 123 babies with paired measurements, the mean difference between the researcher and volunteer measurements was 0.07 cm (95% limits of agreement -0.55 to +0.70) and 7.3% (9/123) of the pairs were outside the 95% limits of agreement. When birth at a health facility is not possible, foot length measurement can identify LBW and PTB in newborns but needs appropriate training for community volunteers and evaluation of its impact on healthcare outcomes.


Gene expression microarray data from human microvascular endothelial cells supplemented with a low concentration of niacin.
The systemic lipid modifying drug, niacin, can directly improve human microvascular endothelial cell angiogenic function under lipotoxic conditions, possibly through activation of niacin receptors "Niacin receptor activation improves human microvascular endothelial cell angiogenic function during lipotoxicity" (Hughes-Large et al. 2014). Here we provide accompanying data collected using Affymetrix GeneChip microarrays to identify changes in gene expression in human microvascular endothelial cells treated with 10 μM niacin. Statistical analyses of robust multi-array average (RMA) values revealed that only 16 genes exhibited greater than 1.3-fold differential expression. Of these 16, only 5 were identified protein coding genes, while 3 of the remaining 11 genes appeared to be small nuclear/nucleolar RNAs. Altered expression of EFCAB4B, NAP1L2, and OR13C8 was confirmed by real time quantitative PCR.


Assisted care as a baseline patient risk characteristic affecting the outcome of transcatheter aortic valve insertion.
Objective measures of frailty have not been well defined as risk factors for a poor outcome after transcatheter aortic valve insertion. We hypothesized that assisted care as a baseline patient characteristic was a simple objective measure of frailty. We reviewed our experience to assess for an association between assisted care and outcome of operation. We retrospectively reviewed the records of 597 patients operated with transcatheter aortic valve insertion from November 2008 through July 2015. The study cohort included patients with a dichotomous baseline characteristic of receiving assisted care (AC group, n = 60, 10.1%) or not receiving assisted care (NC group, n = 537, 89.9%). The endpoints of the study were operative stroke/death and 1-year survival. The age of the patients was 80.6 ± 9.0 years, male sex was present in 349 (58.5%), and STS predicted risk of mortality was 9.2 ± 6.2%. Alternate access was used in 26 (43.3%) patients in the AC care group and in 220 (41.0%) in the NC group (P = 0.724). Operative stroke/death occurred in 4 (6.7%) patients in the AC group and in 25 (4.7%) in the NC group (P = 0.492). Mortality at 1 year in the AC group was 14.8 ± 5.2% and in the NC group was 12.9 ± 1.7%; (P = 0. 250). Assisted care as a baseline patient characteristic does not result in increased operative stroke/death or 1-year mortality in patients following transcatheter aortic valve insertion. Assisted care should not by itself preclude operation. © 2016 Wiley Periodicals, Inc.


A cyclic model of the universe.
We propose a cosmological model in which the universe undergoes an endless sequence of cosmic epochs that begin with a "bang" and end in a "crunch." Temperature and density at the transition remain finite. Instead of having an inflationary epoch, each cycle includes a period of slow accelerated expansion (as recently observed) followed by contraction that produces the homogeneity, flatness, and energy needed to begin the next cycle.


Synchronization of replicons in Ehrlich ascites cells.
Ehrlich ascites cells, in which replication units at the beginning of the S phase started and grew synchronously, were obtained by the following protocol: (1) selection of G1 cells by zonal centrifugation, (2) hypoxia for 12 h, (3) reaeration, (4) addition of cycloheximide (30 microM) within the first minute after reoxygenation. Studies on the effectiveness of the different steps revealed: (i) G1 cells reoxygenated after 12 h of hypoxia traverse two succeeding cell cycles highly synchronously. This was shown by monitoring the thymidine incorporation rate, the thymidine pulse-labeling index, and the mitotic index. (ii) Cycloheximide, like hypoxia, suppresses replicon initiation in Ehrlich ascites cells without interfering with DNA chain growth and DNA maturation. The reversibility of the suppression is less complete than in the case of hypoxia. This was shown by DNA fiber autoradiography and by analyzing the length distribution of pulse- or pulse/pulse-chase-labeled daughter DNA in alkaline sucrose gradients. The alkaline sedimentation patterns of daughter-strand DNA, pulse labeled immediately after the cycloheximide addition at the end of the elaborated protocol and 1 and 2 h later, indicated synchronous initiation and growth of a homogeneous population of DNA molecules to replicon-sized lengths.


Structural studies of the vacuolar H(+)-pyrophosphatase: sequence analysis and identification of the residues modified by fluorescent cyclohexylcarbodiimide and maleimide.
We determined the amino acid residues of the H(+)-translocating inorganic pyrophosphatase (H(+)-PPase) of pumpkin which are covalently labeled by two fluorescent labeling reagents; N-cyclohexyl-N'-[4-(dimethyl amino)-alpha-naphthyl] carbodiimide (NCD) and N-pyrenylmaleimide (NPM). NCD and NPM are fluorescent analogues of N,N-dicycrohexylcarbodiimide and N-ethylmaleimide, respectively, and inactivate H(+)-PPase activity. Excess Mg2+ protected the H(+)-PPase from the inactivation by these reagents. Furthermore, we identified the cDNA clone encoding the pumpkin H(+)-PPase in order to determine the position of labeled residues. The nucleotide sequence of the cDNA clone contains a 2,304 bp open reading frame encoding a polypeptide with 768 amino acids. Chemical sequence analysis of fluorescent peptide fragments revealed that Glu749 located in the C-terminal putative transmembrane alpha-helix was a NCD-labeled residue, and Cys632 was a NPM-labeled residue located in a putative cytosolic domain. The amino acid sequence of the region that includes Glu749 is highly conserved in H(+)-PPases from other plants and it also shows some sequence similarity with the region of the carbodiimide-reactive Glu (or Asp) of F0F1-ATPase c-subunit. The reactive glutamic acids in these proteins are located at the last C-terminal transmembrane alpha-helix. We also found that the H(+)-PPase shows significant amino acid sequence similarity to Kdp-ATPase A chain of E. coli. This similarity between the two different proteins suggest that they have evolved from a common ancestor and may utilize a common basic mechanism for ion transport.


Pure short-chain glycerol fatty acid esters and glycerylic cyclocarbonic fatty acid esters as surface active and antimicrobial coagels protecting surfaces by promoting superhydrophilicity.
Pure glycerol fatty acid esters and glycerylic cyclocarbonic fatty acid esters have an amphiphilic structure, giving these biomolecules a broad range of physico-chemical and biological properties. Physico-chemical properties depend on chain lengths, odd or even carbon numbers on the chain, and glyceryl or cyclocarbonic polar heads. The spectrum of melting-point values for these molecules is large. Surface-activity is very important and through determination of the critical aggregation concentration (CAC), some fatty-acid esters are considered as solvo-surfactant biomolecules. Coupling these self-aggregation and crystallization properties, superhydrophilic surfaces were obtained. An efficient durable water repellent coating of various metallic and polymeric surfaces was allowed. Moreover, these fatty acid esters promoting superhydrophilicity showed biological activity against Gram positive, Gram negative, and yeast-like micro-organisms. Such surfaces coated by self-assembled fatty acid esters in a stable coagel state present a novel solution to surface-contamination risks from pathogen proliferation.


Biosorption Cu (II) by the yeast Saccharomyces cerevisiae.
With the industrial and population advances, the generation of effluents containing heavy metals has grown a lot. In this work, the commercial biomass of the yeast Saccharomyces cerevisiae Perlage® BB were carried out as Cu (II) ion biosorbent. The influence of some variables such as metal concentration, pH range, equilibrium time and biomass concentration were evaluated. The biosorption capacity was measured by adsorption isotherms, with the Langmuir, Freundlich, and Dubinin-Radushkevich (D-R) models. The characterization of the biomass surface were investigated by Dispersive Energy X-Ray Fluorescence Spectrometry (EDX) and Atomic Force Microscopy (AFM). The results showed that the biomass presented good biosorption efficiency. The best fit of the data was obtained with the Langmuir model, detecting the maximum biosorption capacity of 4.73 mg g-1. By the methods used in the characterization of the biomass surface, it was possible to verify the presence of the Cu (II) ion in the yeast.


Hypothalamic fatty acid metabolism mediates the orexigenic action of ghrelin.
Current evidence suggests that hypothalamic fatty acid metabolism may play a role in regulating food intake; however, confirmation that it is a physiologically relevant regulatory system of feeding is still incomplete. Here, we use pharmacological and genetic approaches to demonstrate that the physiological orexigenic response to ghrelin involves specific inhibition of fatty acid biosynthesis induced by AMP-activated protein kinase (AMPK) resulting in decreased hypothalamic levels of malonyl-CoA and increased carnitine palmitoyltransferase 1 (CPT1) activity. In addition, we also demonstrate that fasting downregulates fatty acid synthase (FAS) in a region-specific manner and that this effect is mediated by an AMPK and ghrelin-dependent mechanisms. Thus, decreasing AMPK activity in the ventromedial nucleus of the hypothalamus (VMH) is sufficient to inhibit ghrelin's effects on FAS expression and feeding. Overall, our results indicate that modulation of hypothalamic fatty acid metabolism specifically in the VMH in response to ghrelin is a physiological mechanism that controls feeding.


The Association Between the Rate of Angiotensin-Converting Enzyme Inhibitors and Angiotensin Receptor Blockers Use and the Number of Covid-19 Confirmed Cases and Deaths in the United States: Geospatial Study.
The novel coronavirus SARS-Cov2 uses the angiotensin-converting enzyme 2 (ACE2) receptor as an entry point to the cell. Cardiovascular disease (CVD) is a risk factor for the novel coronavirus disease (Covid-19) with poor outcomes. We hypothesized that the rate of ACE inhibitor (ACEI) and angiotensin receptor blocker (ARB) use is associated with the rate of Covid-19 confirmed cases and deaths. We conducted a geospatial study using publicly available county-level data. The Medicare ACEI and ARB prescription rate was exposure. The Covid-19 confirmed case and death rates were outcomes. Spatial autoregression models were adjusted for the percentage of Black residents, children, residents with at least some college degree, median household income, air quality index, CVD hospitalization rate in Medicare beneficiaries, and CVD death rate in a total county population. After adjustment for confounders, the ACEI use rate did not associate with Covid-19 confirmed case rate (direct county-own effect +0.11 %; 95%CI -0.31 to 0.53; P=0.600, and indirect spillover effect -0.53 %; 95%CI -3.89 to 2.84; P=0.760). The ARB use rate was associated with increased Covid-19 confirmed case rate (direct county-owned effect +0.12 %; 95%CI 0.05-0.19; P=0.002, and indirect spillover effect -0.33 %; 95%CI -2.11 to 1.44; P=0.714). Sensitivity analysis indicated an absence of significant reverse causality bias for analyses with Covid-19 confirmed case rate, but not death rate outcome. Our results highlight the safety of ACEI use for patients with clinical indications for ACEI use. However, an increase in ARB use by 1% was associated with a 0.12 % increase in Covid-19 confirmed cases. The use of ARB, due to known ACE2 upregulation, may facilitate SARS-CoV-2 entry into target cells and increase infectivity. Cluster-randomized controlled trial is warranted to answer the question of whether the replacement of ARB by ACEI may reduce the Covid-19 confirmed case rate.


Feasibility of Laterocaudal Approach in Comparison with Subtarsal Approach in the Management of Orbital Rim Fractures in Indian Population: A Prospective Study.
A balance between surgical access and aesthetics, with lesser post-op sequelae, is inevitable in the management of facial fractures. Even though various approaches are widely practised, further modifications and revisions are encouraged for better outcomes. We performed a prospective cross-sectional study on 30 patients with Zygomatico-Maxillary Complex (ZMC) fracture with inferior orbital rim involvement. Surgical management was performed using subtarsal (Group A) or laterocaudal (Group B) approach, during which the time taken to expose fracture and the ease of access was documented. Scar evaluation was performed using SCAR Scale on the 15th day and after one-month and three-month follow-up, and the post-op complications including ectropion, infections, edema, etc., were documented. There was a significant reduction in the time taken (p < 0.001) in group B than in group A. The ease of access did not demonstrate any significant difference. The SCAR scale score was significantly reduced at 3rd-month follow-up in group B compared to the other group (p = 0.009). Laterocaudal approach demonstrated lesser time-taken and better access to the inferolateral rim, with superior aesthetics and fewer complications. This approach can be employed in clinical practice, especially with inferolateral rim involvement in ZMC fractures.


Usefulness of seismocardiography for the diagnosis of ischemia in patients with coronary artery disease.
Seismocardiography (SCG) is a useful method for the detection of exercise-induced changes in cardiac muscle contractility which may occur during myocardial ischemia. The aim of this study was to compare the diagnostic accuracy of SCG with electrocardiographic exercise test (ETT) for diagnosis of ischemia in patients with angiographically proved coronary artery disease (CAD). Seventy-seven male patients with CAD without myocardial infarction (MI), mean age 51+/-9 years, were subjected to SCG and ETT. A gender-matched control group consisted of 30 healthy volunteers aged 34+/-7 years. SCG was done simultaneously with resting supine 12-lead electrocardiography before and immediately after a symptom-limited ETT. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) of SCG were compared with ETT. Moreover, the diagnostic accuracy of both the methods was compared, with coronary angiography being the reference for the analysis. SCG was more sensitive (61.1% vs 44.2%, P<0.05) and accurate (70% vs 61%, P<0.05) method for detecting ischemia caused by coronary stenosis>or=50%, at least in one coronary artery compared to the ETT. However, ETT had better specificity than SCG (82.4% vs 76%, P<0.05). The PPV and NPV of SCG were significantly better than those obtained with ETT (77.9% vs 76%, P<0.05 and 63.4% vs 53.8%, P<0.05, respectively). Moreover, the concordant results of SCG and ETT improved the diagnostic accuracy of both methods. SCG appeared to be more sensitive for detecting ischemia caused by more than>or=50% stenosis of the main coronary artery compared to an electrocardiographic stress test. SCG was a useful ETT adjunct for selecting patients requiring coronary angiography.


Possible subtypes of ATP receptor producing contraction of rat vas deferens, revealed by cross-desensitisation.
1. Adenosine triphosphate (ATP), alpha beta-methylene ATP and the dinucleotides P1P4diadenosine tetraphosphate (AP4A) and P1P5-diadenosine pentaphosphate (AP5A) have been applied to preparations of the rat vas deferens in vitro in preliminary experiments designed to assess the feasibility of using such nucleotides to detect subtypes of the ATP receptor causing smooth muscle contractions. 2. Desensitisation to the dinucleotides induced a change in the profile of the response to ATP with the preferential loss of an early transient component. 3. Desensitisation to low concentrations of alpha beta-methylene ATP produced a greater loss of the later, slower component of contraction to ATP. 4. It is suggested that these components may therefore involve pharmacologically distinguishable nucleotide receptors.


Chimerical anterolateral thigh flap for plantar reconstruction.
The purpose of this study is to evaluate the results of reconstruction and rehabilitation of patients with plantar defects by using a chimerical flap of muscle and skin from anterolateral thigh. Twenty-five patients with plantar defects were reconstructed with a chimerical anterolateral thigh (ALT) flap, composed by a vastus lateralis muscle segment and a thinned skin island. Neurorrhaphy between lateral femoral cutaneous nerve and calcaneal nerve was performed in 7 patients. Evaluation of flap contour and stability and patient ambulation was performed 6 and 12 months after surgery. Evaluation of cutaneous sensiblity of ALT flap and contralateral thigh was performed 12 months after surgery using Pressure Specified Sensory Device™ (PSSD™). Flap viability was complete in 23 patients and 2 patients had complications with partial flap loss of its cutaneous component. Six months postoperatively, flap contour, and stability was considered good in 19 and 21 patients respectively, and all 25 patients presented good ambulation. Twelve months postoperatively, all 25 patients presented good flap contour and stability, as well as good ambulation. All 7 flaps undergoing to reinnervation partially recovered cutaneous sensibility in comparison to donor site (contralateral thigh). Cutaneous tactile thresholds (g/mm(2) ) of static one-point test and moving one-point test from the ALT flap and the contralateral thigh presented statistically significant differences, for both comparisons (P = 0.009, P = 0.002). This flap is suitable for reconstruction of plantar defects, with good flap contour and stability, proper patient ambulation and low complication rates.


PDE3A gene screening improves diagnostics for patients with Bilginturan syndrome (hypertension and brachydactyly syndrome).
Autosomal-dominant hypertension and brachydactyly syndrome (HTNB; Bilginturan syndrome) is known to cause stroke before age 50 when untreated. We report a novel PDE3A gene mutation in a mother and daughter affected with dominant brachydactyly of the hands and feet, a short stature, and hypertension. The hypertension was medically responsive to anti-hypertensive treatment. The 3-bp deletion in the PDE3A gene presented de novo in the mother. Here, we expand the list of PDE3A mutations identified in Bilginturan syndrome and emphasize the importance of standardized genetic testing of HTNB patients to improve diagnostics at an early age. We recommend extended phenotyping in patients with brachydactyly, a short stature or hypertension in clinical practice.