Utilizing Closed Incisional Negative Pressure Therapy Reduces Peripheral Bypass Infection Rates Without Increasing Costs.
We evaluated outcomes of closed incisional negative pressure therapy (ciNPT) on surgical site infection (SSI) rates in lower extremity bypass patients. We sought to determine whether or not the routine use of ciNPT is a cost-effective measure. During a period from May 2018 to August 2018, our institution transitioned to the routine use of ciNPT for re-vascularization procedures. We retrospectively reviewed our outcomes before and after the initiation of ciNPT. Group A included patients from September 2017 to April 2018 without ciNPT and Group B included patients from September 2018 to April 2019 with ciNPT. Chi-squared analysis was performed and the p value was set at <0.05 to obtain statistical significance. Cost analysis was separately performed utilizing hospital metrics. There were a total of 102 patients in Group A and 113 patients in Group B. There was no difference in demographic information between the two groups. The overall SSI rate for Group A was 11.8% (12/102). Group B had an overall SSI rate of 3.5% (4/113; p=0.02). Deep infection rate for Group A was 7% (7/102) and for Group B was 1% (1/113; p=0.01). Cost analysis demonstrated a minimum of $62,000 in infection-related cost savings between both groups. ciNPT has had a profound effect on our practice and has resulted in a decrease in both deep and superficial infections. This has led to a significant cost-effective measure for our institution. We now routinely use ciNPT on all lower extremity bypass patients.


Molecular and genetic prognostic factors of prostate cancer.
Prostate cancer is the most commonly diagnosed cancer in Western males, responsible for 3% of all deaths in men over 55 years of age and second only to lung cancer as a cause of cancer death. Biomarkers have become an important diagnostic tool in prostate cancer. The discovery of the serum marker prostate-specific antigen (PSA) significantly facilitated the detection and management of prostate cancer. As we enter into the post-genomics era, novel biomarkers of prostate cancer of therapeutic significance will invariably emerge. Here we review a series of existing and emerging molecular-based prognostic markers particularly with radiotherapy.


Magnetic resonance imaging of noncommunicating hydrocephalus.
The purpose of this review is to describe the magnetic resonance imaging (MRI) findings in patients with noncommunicating hydrocephalus (NCH). We describe the technical aspects of MR sequences that are of use for the imaging of cerebrospinal fluid (CSF). Both the diagnosis of NCH and the follow-up after endoscopic third ventriculostomy (ETV) are discussed through clinical cases. NCH is related to an obstacle hindering the CSF pathways. The combined use of anatomical and flow-sensitive MR sequences may help to characterize hydrocephalus. An imaging protocol is described at 1.5-T. The first step of the diagnostic work-up consists of the detection of an obstacle hindering CSF flow pathways using both constructive interference in steady state and flow-sensitive MR sequences. CSF volumetry and quantification of ventricular wall movement may also improve the diagnosis and follow-up of patients with NCH.


In vitro efficacy of the combination of ciprofloxacin and cefotaxime against Vibrio vulnificus.
We performed time-kill studies of antimicrobial combinations that included minocycline, cefotaxime, and ciprofloxacin with Vibrio vulnificus ATCC 27562. Cefotaxime-plus-ciprofloxacin combinations acted synergistically against V. vulnificus in vitro, and this combination regimen can be a good choice as the empirical treatment for suspected necrotizing fasciitis due to V. vulnificus.


Esophageal diverticulum associated with Aerococcus viridans infection in a loggerhead sea turtle (Caretta caretta).
A juvenile loggerhead sea turtle (Caretta caretta) stranded in Gran Canaria, Spain was necropsied. The turtle was underweight, had sunken eyes, and small amounts of crude oil were in the oral cavity. The most significant lesion was a large esophageal diverticulum at the junction of the esophagus and stomach. The diverticulum was full of gas and green mucoid fluid and a diffuse thick yellow fibrinonecrotic membrane covered the mucosa. The lumen of the diverticulum also contained moderate numbers of cephalopods, crustaceans, and anthropogenic debris including crude oil balls, plastics, and fishing lines. Histologically there was a severe diffuse fibrinonecrotic esophagitis. Aerococcus viridans was isolated from the diverticulum. This is the first report of an esophageal diverticulum in a sea turtle. Although A. viridans is a known pathogen of lobsters and fishes, there are no reports of A. viridans infection in sea turtles.


Results of treatment in patients with end-stage renal disease: a multivariate analysis of risk factors and survival in 341 successive patients.
Factors that affect survival in patients with end-stage renal disease (ESRD) are still only partially understood. We report an analysis on a cohort of 341 successive patients who started ESRD treatment in one institution. Survival was calculated from the start of ESRD treatment, whether initial treatment was by dialysis (335 patients) or transplantation (6 patients). Analysis of factors affecting survival was done using the Cox multivariate hazard analysis. Relative risks were calculated for several demographic factors, the primary renal disease, the presence or absence of various high-risk co-morbid factors, and for a first transplant from a living-related or cadaver donor. Older patients, patients with no prior health insurance, those with diabetic nephropathy, and those with small kidneys of unknown cause had statistically significantly higher risks of dying. The risk of dying decreased by year of start of ESRD treatment. Living related donor transplantation was associated with a decreased hazard to age 45 years and older; whereas cadaver donor transplantation was associated with a hazard that increased with age and was significantly increased by age 45.


Fluorine-18-Labeled Antibody Ligands for PET Imaging of Amyloid-β in Brain.
Antibodies are attractive as radioligands due to their outstanding specificity and high affinity, but their inability to cross the blood-brain barrier (BBB) limits their use for CNS targets. To enhance brain distribution, amyloid-β (Aβ) antibodies were fused to a transferrin receptor (TfR) antibody fragment, enabling receptor mediated transport across the BBB. The aim of this study was to label these bispecific antibodies with fluorine-18 and use them for Aβ PET imaging. Bispecific antibody ligands RmAb158-scFv8D3 and Tribody A2, both targeting Aβ and TfR, were functionalized with trans-cyclooctene (TCO) groups and conjugated with 18F-labeled tetrazines through an inverse electron demand Diels-Alder reaction performed at ambient temperature. 18F-labeling did not affect antibody binding in vitro, and initial brain uptake was high. Conjugates with the first tetrazine variant ([18F]T1) displayed high uptake in bone, indicating extensive defluorination, a problem that was resolved with the second and third tetrazine variants ([18F]T2 and [18F]T3). Although the antibody ligands' half-life in blood was too long to optimally match the physical half-life of fluorine-18 (t1/2 = 110 min), [18F]T3-Tribody A2 PET seemed to discriminate transgenic mice (tg-ArcSwe) with Aβ deposits from wild-type mice 12 h after injection. This study demonstrates that 18F-labeling of bispecific, brain penetrating antibodies is feasible and, with further optimization, could be used for CNS PET imaging.


Degradation mechanism and the toxicity assessment in TiO2 photocatalysis and photolysis of parathion.
The photocatalytic degradation of methyl parathion was carried out using a circulating TiO2/UV reactor. The experimental results showed that parathion was more effectively degraded in the photocatalytic condition than the photolysis and TiO2-only condition. With photocatalysis, 10mg/l parathion was completely degraded within 60 min with a TOC decrease exceeding 90% after 150 min. The main ionic byproducts during photocatalysis were measured. The nitrogen from parathion was recovered mainly as NO3-, NO2- and NH4+, 80% of the sulfur as SO4(2-), and less than 5% of the phosphorus as PO4(3-). The organic intermediates 4-nitrophenol and paraoxon were also identified, and these were further degraded. Two different bioassays (Vibrio fischeri and Daphnia magna) were used to test the acute toxicity of solutions treated by photocatalysis and photolysis. A Microtox test using V. fischeri showed that the toxicity, expressed as the relative toxicity (%), was reduced almost completely after 90 min under photocatalysis, whereas only an 83% reduction was achieved with photolysis alone. Another toxicity test using D. magna also showed that the relative toxicity disappeared after 90 min under photocatalysis, whereas there was a 65% reduction in relative toxicity with photolysis alone. The pattern of toxicity reduction parallels the decrease in parathion and TOC concentrations.


Abnormal Presentation of Hypoxic Ischemic Encephalopathy Attributed to Polysubstance Exposure.
BACKGROUND With the increasing prevalence of substance use in pregnancy, the rates of neonatal abstinence syndrome (NAS) are dramatically increasing. There is little information on the use of multiple substances in adults, even less so of polysubstance abuse during pregnancy and the consequences for the fetus as well as the mother. CASE REPORT A newborn male born at 35 weeks presented post-delivery with hips bilaterally dislocated and hyperflexed. The patient's legs fully extended and their shoulders were bilaterally mid-flexed with arms fully extended. This neonate was also reported to have bilateral hearing and vision loss as well as NAS symptoms of high-pitched crying and respiratory distress. During pregnancy the mother in this case study admitted to using buprenorphine, benzodiazepines, gabapentin, and heroin. The consequences of using this combination has not been well studied in pregnancy. CONCLUSIONS The presented case had severe complications, likely due to maternal polysubstance use and poor prenatal care in pregnancy. Clonidine was used to control the NAS symptoms, ranitidine was used to treat the gastroesophageal reflux, and glycopyrronium bromide was used for the neonate's excessive secretions. After delivery, the patient was placed on a nasal noninvasive cannula for respiratory distress and was transferred to a different hospital for treatment of the more serious comorbid conditions.


Peculiar effects of muscarinic M1, M2, and M3 receptor blockers on cardiac chronotropic function in neonatal rats.
The effects of muscarinic M(1), M(2), and M(3) cholinergic receptor blockade on the regulation of chronotropic function of the heart were studied in vivo in 7-day-old rat pups. Intravenous injection of M(2) receptor blocker gallamine produced no changes in cardiac chronotropy. In contrast, M(1) receptor blocker pirenzepine and M(3) receptor blocker 4DAMP provoked bradycardia. These data attest to the involvement of M(1) and especially M(3) cholinergic receptors in the regulation of cardiac chronotropy in rat pups, which confirms the view on pronounced species-specific and age-related peculiarities in the heart control mechanisms.


American College of Radiology-American Brachytherapy Society practice parameter for electronically generated low-energy radiation sources.
This collaborative practice parameter technical standard has been created between the American College of Radiology and American Brachytherapy Society to guide the usage of electronically generated low energy radiation sources (ELSs). It refers to the use of electronic X-ray sources with peak voltages up to 120 kVp to deliver therapeutic radiation therapy. The parameter provides a guideline for utilizing ELS, including patient selection and consent, treatment planning, and delivery processes. The parameter reviews the published clinical data with regard to ELS results in skin, breast, and other cancers. This technical standard recommends appropriate qualifications of the involved personnel. The parameter reviews the technical issues relating to equipment specifications as well as patient and personnel safety. Regarding suggestions for educational programs with regard to this parameter,it is suggested that the training level for clinicians be equivalent to that for other radiation therapies. It also suggests that ELS must be done using the same standards of quality and safety as those in place for other forms of radiation therapy.


Rebound in Measures of Disease Activity and Symptoms in Corrona Registry Patients with Psoriatic Arthritis Who Discontinue Tumor Necrosis Factor Inhibitor Therapy after Achieving Low Disease Activity.
Rebound may occur in patients with psoriatic arthritis (PsA) who discontinue TNF inhibitor (TNFi) therapy in low disease activity (LDA). Using physician and patient reports, we quantified rebound following TNFi discontinuation [defined as Clinical Disease Activity Index (CDAI) score > 10 or TNFi restart] and time to rebound in adults with PsA in LDA (CDAI score ≤ 10) at TNFi discontinuation. Rebound occurred in 73% (69/94) of patients soon after discontinuation (median time to rebound 8.0 mos, 95% CI 6.0-12.0). Rebound occurred frequently in patients with PsA after TNFi discontinuation. TNFi discontinuation after achieving LDA should be carefully considered.


Exploring Workforce Participation Patterns and Chronic Diseases Among Middle-Aged Australian Men and Women Over the Life Course.
This study identified associations between chronic diseases (diabetes, asthma, depression, and arthritis) and workforce participation patterns with a gendered perspective. We used data from 1,261 middle-aged participants of the Australian Life Histories and Health (LHH) Survey, aged 60 to 64 years in 2011. Latent class analysis identified dominant workforce patterns and associations between chronic diseases and these patterns were explored by multinomial regression models. Diabetes, asthma, depression, and arthritis were less prevalent in men and women in class "mostly full-time work," compared with other workforce patterns. The odds of "mostly full-time work" were lower for men reporting depression or arthritis, whereas among women, depression was associated with "increasing part-time work" after adjusting early and adult life factors. The results strengthen the importance of gender focused policies aimed to promote and preserve health of young and middle-aged workers, and creating supportive environment for those with chronic health issues over the life course.


Selected estrogen receptor 1 and androgen receptor gene polymorphisms in relation to risk of breast cancer and fibrocystic breast conditions among Chinese women.
Polymorphisms in sex hormone receptor-encoding genes may alter the activity of sex hormone receptors and thereby affect susceptibility to breast cancer and related outcomes. In a case-control study of women from Shanghai, China, we examined the risk of breast cancer and fibrocystic breast conditions associated with the ESR1 PvuII (rs2234693) and XbaI (rs9340799) and AR CAG repeat ((CAG)(n)) and GGC repeat ((GGC)(n)) polymorphisms among 614 women with breast cancer, 467 women with fibrocystic conditions, and 879 women without breast disease. We also evaluated whether risk differed by the presence/absence of proliferative changes (in the extratumoral epithelium or fibrocystic lesion), menopausal status, or body mass index (BMI). Age-adjusted odds ratios (ORs) and 95% confidence intervals (95% CI) were calculated using logistic regression. Only associations with AR (CAG)(n) and (GGC)(n) genotypes were detected. Allocating AR (CAG)(n) genotypes into six categories, with the (CAG)(22-24)/(CAG)(22-24) genotype category designated as the reference group, the (CAG)(>24)/(CAG)(>24) genotype category was associated with an increased risk of fibrocystic breast conditions (OR, 1.8; 95% CI, 1.1-3.0). Relative to the AR (GGC)(17)/(GGC)(17) genotype, the (GGC)(17)/(GGC)(14) genotype was associated with elevated risks of incident breast cancer (OR, 2.6; 95% CI, 1.3-5.4) and fibrocystic conditions (OR, 2.3; 95% CI, 1.1-4.5). Results did not differ according to proliferation status, menopausal status, or BMI. Although these data lend support for a link between AR variation and breast disease development, given the low frequency of the putative risk-conferring genotypes and other constraints, further confirmation of our results is needed.


Short rib-polydactyly syndrome, Majewski type, in two male siblings.
Two male siblings with a short rib-polydactyly syndrome, Majewski type (Majewski syndrome), are reported. To our knowledge, the present report is the first one of siblings each confirmed to be affected by Majewski syndrome, and it may therefore contribute to establishing the mode of inheritance.


Dynamic limitation of coronary vasodilator reserve in patients with dilated cardiomyopathy and chest pain.
Twenty-six patients with dilated cardiomyopathy and angiographically normal coronary arteries, 12 of whom gave a history of anginal chest pain, underwent noninvasive and invasive hemodynamic study. During treadmill exercise testing, patients with a history of angina demonstrated worse effort tolerance (7.4 +/- 4.9 versus 13.6 +/- 5.1 minutes, p less than 0.005) and a lower end-exercise systolic blood pressure-heart rate product (17.9 +/- 3.4 versus 23.6 +/- 4.9 mm Hg.beats/min x 10(3), p less than 0.005) compared with patients without a history of angina. During rapid atrial pacing after ergonovine, 0.15 mg intravenously, 11 of the 12 patients with a history of angina experienced their typical chest pain, in contrast to only 1 of 12 patients without a history of angina. The angina group, compared with the nonangina group, had significantly lower great cardiac vein flow (118 +/- 24 versus 160 +/- 43 ml/min, p less than 0.01), and higher coronary resistance (0.87 +/- 0.21 versus 0.66 +/- 0.25 mm Hg.min/ml, p less than 0.05), significant widening of the arterial--great cardiac vein oxygen difference and a significant fall in cardiac index during pacing. Further, ergonovine resulted in higher coronary resistance during pacing in the angina group compared with pacing alone (+0.16 +/- 0.16 mm Hg min/ml, p less than 0.01), in the absence of significant reduction in epicardial coronary artery luminal diameter. After dipyridamole, 0.5 to 0.75 mg/kg intravenously, to 21 patients, the 7 patients with a history of angina had significantly lower flow (149 +/- 37 versus 218 +/- 73 ml/min, p less than 0.05) and higher coronary resistance (0.59 +/- 0.09 versus 0.43 +/- 0.17 mm Hg.min/ml, p less than 0.05) than did the nonangina group. It is concluded that patients with dilated cardiomyopathy and chest pain unrelated to epicardial coronary artery disease exhibit impaired vasodilator responses to both metabolic and pharmacologic stimuli, and an increased sensitivity to the vasoconstrictor effects of ergonovine. Whether these findings are of etiologic or long-term prognostic significance is unknown.


Proteinuria due to suboptimal hydration with high-dose methotrexate therapy.
One of the major complications after high-dose methotrexate (HDMTX) infusions is renal damage. We investigated the occurrence of proteinuria after HDMTX administration in children with pediatric malignancies (acute lymphoid leukaemia, osteosarcoma Burkitt's lymphoma). In the period 1989-1990 we gave 52 HDMTX courses to 24 children. During this period, prehydration and extra urinary alkalisation were performed only if the urinary specific gravity was over 1010 or if the urinary pH fell below 7. Using this schedule the mean values obtained for protein extraction were: before the therapy, 0.12 +/- 0.03 g/m2; on day 1 after MTX treatment, 0.38 +/- 0.06 g/m2; and on day 2 after the MTX infusion, 0.39 +/- 0.11 g/m2 (P < 0.01). A significant increase in proteinuria (> 0.2 g/m2 post- vs pretreatment) was detectable in 54% of the patients. In the period 1991-1992 we modified the hydration-alkalisation schedule to include i.v. prehydration for 18-24 h at 3 l/m2/day with a 0.45% NaCl-5% glucose solution along with sodium bicarbonate and posthydration for 72 h with the same solution. On this protocol the mean values determined for the urinary protein content were all in the normal range (pretreatment, 0.03 g/m2/day; day 1, 0.05 g/m2/day; and day 2, 0.08 g/m2/day). These findings were significantly different from the previous results (P < 0.05).


Bayesian data analysis in the phonetic sciences: A tutorial introduction.
This tutorial analyzes voice onset time (VOT) data from Dongbei (Northeastern) Mandarin Chinese and North American English to demonstrate how Bayesian linear mixed models can be fit using the programming language Stan via the R package brms. Through this case study, we demonstrate some of the advantages of the Bayesian framework: researchers can (i) flexibly define the underlying process that they believe to have generated the data; (ii) obtain direct information regarding the uncertainty about the parameter that relates the data to the theoretical question being studied; and (iii) incorporate prior knowledge into the analysis. Getting started with Bayesian modeling can be challenging, especially when one is trying to model one's own (often unique) data. It is difficult to see how one can apply general principles described in textbooks to one's own specific research problem. We address this barrier to using Bayesian methods by providing three detailed examples, with source code to allow easy reproducibility. The examples presented are intended to give the reader a flavor of the process of model-fitting; suggestions for further study are also provided. All data and code are available from: https://osf.io/g4zpv.


[Effects of handling and processing on deoxynivalenol and zearalenone content of cereals and cereal products].
Since national limits have been introduced for the content of DON and ZEA in cereals and cereal products designated for human consumption, it is highly important to understand how these toxins are distributed during sorting, cleaning and further processing to bakery products and pasta. Cereals from several crops were analysed before and after sorting and cleaning. After milling, flours, breads, semolinas, pastas and others were analysed. The results show that that the distribution of DON and ZEA was different. ZEA was more effectively removed than DON. The efficacy of the various processes varied markedly from one lot to the other.


The First 24 Hours: Burn Shock Resuscitation and Early Complications.
Resuscitation is required for the management of patients with severe thermal injury. Some of the initial pathophysiologic events following burn injury include an exaggerated inflammatory state, injury to the endothelium, and increased capillary permeability, which all culminate in shock. Understanding these processes is critical to the effective management of patients with burn injuries. Formulas predicting fluid requirements during burn resuscitation have evolved over the past century in response to clinical experience and research efforts. Modern resuscitation features individualized fluid titration and monitoring along with colloid-based adjuncts. Despite these developments, complications from over-resuscitation still occur.


Effect of finishing diets on Escherichia coli populations and prevalence of enterohaemorrhagic E. coli virulence genes in cattle faeces.
To determine the effect of different carbohydrate-based finishing diets on fermentation characteristics and the shedding of Escherichia coli and enterohaemorrhagic E. coli (EHEC) virulence genes in cattle faeces. The size of faecal E. coli populations and fermentation characteristics were ascertained in three experiments where cattle were maintained on a range of finishing diets including high grain, roughage, and roughage + molasses (50%) diets. Increased E. coli numbers, decreased pH and enhanced butyrate and lactate fermentation pathways were associated with grain diets, whereas roughage and roughage + molasses diets resulted in decreased concentrations of ehxA, eaeA and stx(1) genes, this trend remaining at lairage. In one experiment, faecal E. coli numbers were significantly lower in animals fed roughage and roughage + molasses, than animals fed grain (4.5, 5.2 and 6.3 mean log10 g(-1) digesta respectively). In a second experiment, faecal E. coli numbers were 2 log lower in the roughage and roughage + molasses diets compared with grain-fed animals prior to lairage (5.6, 5.5 and 7.9 mean log10 g(-1) digesta respectively) this difference increasing to 2.5 log at lairage. The type of dietary carbohydrate has a significant effect on E. coli numbers and concentration of EHEC virulence genes in faeces of cattle. The study provides a better understanding of the impact finishing diet and commercial lairage management practices may have on the shedding of E. coli and EHEC virulence factors, thus reducing the risk of carcass contamination by EHEC.


Autophagy regulation in macrophages and neutrophils.
Autophagy is a conserved proteolytic mechanism that degrades cytoplasmic material including cell organelles. Accumulating evidence exists that autophagy also plays a major role in immunity and inflammation. Specifically, it appears that autophagy protects against infections and inflammation. Here, we review recent work performed in macrophages and neutrophils, which both represent critical phagocytes in mammalians.


ANCA-associated vasculitis in systemic sclerosis report of 3 cases.
The aim of the study was to describe the occurrence of anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) in systemic sclerosis (SSc) patients. SSc patients who developed biopsy-proven AAV were identified. Their clinical manifestations, autoantibodies, presentation with vasculitis, treatment and outcome were described and compared with previously reported patients with these two conditions. Of 985 patients, 3 were identified. All patients had interstitial lung disease, and all presented with acute renal failure, proteinuria and hematuria, and were P-ANCA- and anti-Scl-70-positive. One required hemodialysis. Two were hypertensive; additionally, one patient had sinusitis, and another had monoarthritis and a macular rash. All were treated with high-dose corticosteroids and responded to therapy and attained remission at 6 months. At 1 year, one patient died of pneumonia. ANCA-associated vasculitis is a rare but serious finding in SSc patients. Positive anti-Scl-70 antibody is found commonly in these patients. Different treatment modalities are effective. Serious infections can complicate therapy and lead to death.


Assessing healthcare workers' knowledge, emotions and perceived institutional preparedness about COVID-19 pandemic at Saudi hospitals in the early phase of the pandemic.
Background: Coronavirus disease 2019 (COVID-19) pandemic extended to reach most countries in the globe during few months. Preparedness of healthcare institutions and healthcare workers (HCWs) are crucial for applying effective prevention and control measures. This study aimed to assess HCWs knowledge, emotions and perception of preparedness of their institutions towards COVID-19 pandemic. Design: A cross-sectional, web-based survey was conducted among hospital HCWs in Saudi Arabia during April 27, 2020 to May 03, 2020. Results: Overall, 1004 completed responses were received. The majority were females (78.8), nurses (84.9%) at middle age 25-39 years (71.8%). Among participants, 95.5% reported receiving training on safely use of personal protective equipment (PPE) and 94.9% did fit the test for N95 respirator. The participants possessed a fair knowledge about COVID-19 disease with a mean knowledge score 6. 61±1.35 points on a scale of 10 points. Most participants (88.7%) were committed to continue work as a professional and ethical duty, however, 27.1% of them scored high on a negative emotional impact scale. Participants appreciated most aspects of institutional preparedness for COVID-19 pandemic; however, they were concerned with the continuous PPE supply. Factors that independently associated with good knowledge and negative emotional response were determined using multivariate logistic regression analysis. Conclusions: Findings revealed fair knowledge about COVID-19 pandemic among HCWs in Saudi hospitals. Concerns and worries were expressed regard working with the highly infectious COVID-19 patients. Participants, appreciated most aspects of institutional preparedness, however they were concerned about the continuous availability and supply of PPE.


Microbial production and industrial applications of keratinases: an overview.
Massive production of keratinaceous byproducts in the form of agricultural and industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This review summarizes the potential utility of some bacterial and fungal species for the production of keratinase using a variety of keratinaceous wastes as growth substrates. The application of microbial keratinases in waste management; animal feed, detergent, and fertilizer manufacturing; and leather, cosmetic, and pharmaceutical industries is also abridged in this review.


Multicenter clinical trial on dental implants survival rate: a FDS76® study.
Implantology, thanks to its predictability in oral rehabilitations, has become a widespread method for rehabilitating edentulous patients, offering excellent patient satisfaction. Improving the quality of oral health, especially if rehabilitation involves minimal short and long-term complications, leads to an improvement in the quality of life of our patients. This clinical study was conducted to evaluate the short and long-term complications (up to 18 months) of implant-prosthetic rehabilitations performed with a new dental implant (FDS76®, Italy). The results showed a limited number of complications or implant failures. Knowing the latter could certainly help improve both surgical and dental materials performance.


[Comparative analysis of Ca(2+)-signalling in brown preadipocytes of ground squirrel Spermophillus undulatus and mouse].
Analysis of the slow Ca(2+)-responses of brown preadipocytes of ground squirrel Spermophillus undulatus and mouse was carried out. The mouse brown preadipocytes demonstrated low but prominent responses to noradrenalin with the maximum at 3 and 10 microM being the less effective. The ground squirrel brown preadipocytes practically did not practically respond to 10 nM-10 microM, whereas 30-600 microM noradrenalin was able to raise intracellular [Ca2+]i up to 600 nM with 300 microM agonist being the most effective. Stimulation of the plasma membrane Ca(2+)-channels with thimerosal showed considerable reduction of the calcium entry system in the cell precursors of both species comparing with their mature adipocytes. Intracellular calcium stores liberated in preadipocytes of both species by tapsigargin and ionomycin in Ca(2+)-free medium were insignificant, and capacitative Ca(2+)-entry in response to the cellular Ca(2+)-stores depletion was completely absent in Ca(2+)-containing medium. The Ca(2+)-responses of the ground squirrel brown preadipocytes were independent on physiological state of the animals and annual seasons. Preadipocytes of both species showed the same dose-response curves for the Ca(2+)-raise under thimerosal, and the mouse had two-fold higher kinetic constants for the Ca2+ ions entry. The ground squirrel brown adipocytes responded to ionomycin with approximately 25% higher increase in [Ca2+]i and the entry of the ions had 7-10-fold higher kinetic constants for this process. Kinetic constants for the [Ca2+]i raise in mouse preadipocytes were independent of ionomycin concentration, whereas in the ground squirrel brown preadipocytes the constant linearly increased with the ionophore concentration. It is suggested that the found difference in the function of Ca(2+)-signalling in preadipocytes of two species, which becomes apparent in the presence of ionomycin, might be responsible for the observed difference in the noradrenalin induced cellular Ca(2+)-responses as well.


Knowledge, Attitudes, and Intentions towards Human Papillomavirus Vaccination among Nursing Students in Spain.
Human papillomavirus (HPV), which is linked to specific types of cancer, can be prevented by vaccination. This study aimed to determine the knowledge and attitudes of nursing students about HPV and its vaccine as well as their intentions towards personal vaccination. A total of 536 Spanish nursing students were invited to complete the Spanish version of the questionnaire "Knowledge, attitudes and intentions towards HPV". Overall, 367 surveys were completed (68.4% response rate). Data analysis included the calculation of three scores: the knowledge score, categorized into low (<33%), moderate (33%-66%), and good knowledge (>66%); the attitude score, sorted into positive (<2.5), neutral (2.5-3.5), and negative attitude (>3.5); and the intention score, categorized into not favorable (<4), neutral (4-7), and favorable intention (>7). Knowledge about HPV and its vaccine was moderate (54.34 ± 0.9%), and the attitude towards vaccination was positive (2.34 ± 0.03). The intention towards personal vaccination increased significantly after completing the questionnaire (before: 4.14 ± 0.27, after: 6.02 ± 0.28; p < 0.001). The present study highlights the need of training future nurses about HPV and its vaccine, considering the important role it plays in the prevention of sexually transmitted diseases.


Differential regulation of distinct Vps34 complexes by AMPK in nutrient stress and autophagy.
Autophagy is a stress response protecting cells from unfavorable conditions, such as nutrient starvation. The class III phosphatidylinositol-3 kinase, Vps34, forms multiple complexes and regulates both intracellular vesicle trafficking and autophagy induction. Here, we show that AMPK plays a key role in regulating different Vps34 complexes. AMPK inhibits the nonautophagy Vps34 complex by phosphorylating T163/S165 in Vps34 and therefore suppresses overall PI(3)P production and protects cells from starvation. In parallel, AMPK activates the proautophagy Vps34 complex by phosphorylating S91/S94 in Beclin1 to induce autophagy. Atg14L, an autophagy-essential gene present only in the proautophagy Vps34 complex, inhibits Vps34 phosphorylation but increases Beclin1 phosphorylation by AMPK. As such, Atg14L dictates the differential regulation (either inhibition or activation) of different Vps34 complexes in response to glucose starvation. Our study reveals an intricate molecular regulation of Vps34 complexes by AMPK in nutrient stress response and autophagy.


Inference of alternative splicing from tiling array data.
Alternative splicing (AS) is an important mechanism implicated in eukaryotic gene expression, whereby exon segments of precursor-mRNA transcripts are joined together in different arrangements corresponding to diverse isoforms of mature mRNA. Accumulating evidence suggests that in many instances this process is specifically regulated and contributes to the structural and functional diversification of tissues and cell types. Furthermore, several studies support the view that environmental stresses dramatically impact on AS and reported the presence of novel transcript isoforms in response to biotic or abiotic stresses. Since specific regulation of AS in plants is a largely unexplored field of research, large-scale approaches aimed at monitoring AS on a genome-wide level are of increasing importance to gain insights into tissue-specific splicing regulation and to study the effects of changed environmental conditions on pre-mRNA splicing.Here, we describe the concepts of a traditional statistical approach, and a more recently developed machine learning-based method for AS detection from tiling arrays. The here presented approaches were employed for the detection and profiling of AS events in the model plant A. thaliana, and applied to a large dataset comprising transcriptomic expression data from 11 tissues and 13 stress conditions.


Profiling motility signal-specific genes in primary human keratinocytes.
Regulation of human keratinocyte (HK) migration is critical for skin wound healing. Profiling HK migration-specific genes could help us gain a comprehensive understanding of the process. The main challenge is to separate genes that are unrelated to migration, but simultaneously induced by the same growth factor. In this study, we took advantage of a unique response of HKs to transforming growth factor-beta (TGF-beta), which inhibits proliferation but not migration of HKs, to suppress selectively the proliferation-related genes. Furthermore we stimulated HKs independently with TGF-alpha or insulin and identified the common genes and eliminated TGF-alpha- or insulin-specific genes. Under these conditions, we obtained profiles of the immediate-early genes (IEGs, at 30 minutes), early genes (EGs, at 60 minutes), and delayed-early genes (DEGs, at 120 minutes) by microarray analyses, followed by quantitative real-time reverse transcription-PCR (QRT-PCR) validation and functional characterization by RNA interference (RNAi). Our results revealed the following: (1) 25 upregulated and 1 downregulated IEGs; (2) 58 upregulated and 15 downregulated EGs, and (3) 13 upregulated and 3 downregulated DEGs in both TGF-alpha- and insulin-stimulated HKs. Three genes, all encoding secreted molecules, were investigated in HK migration. These cell motility-specific gene profiles may prove useful to skin wound healing.


Recruitment of Mediator Complex by Cell Type and Stage-Specific Factors Required for Tissue-Specific TAF Dependent Gene Activation in an Adult Stem Cell Lineage.
Onset of terminal differentiation in adult stem cell lineages is commonly marked by robust activation of new transcriptional programs required to make the appropriate differentiated cell type(s). In the Drosophila male germ line stem cell lineage, the switch from proliferating spermatogonia to spermatocyte is accompanied by one of the most dramatic transcriptional changes in the fly, as over 1000 new transcripts turn on in preparation for meiosis and spermatid differentiation. Here we show that function of the coactivator complex Mediator is required for activation of hundreds of new transcripts in the spermatocyte program. Mediator appears to act in a sequential hierarchy, with the testis activating Complex (tMAC), a cell type specific form of the Mip/dREAM general repressor, required to recruit Mediator subunits to the chromatin, and Mediator function required to recruit the testis TAFs (tTAFs), spermatocyte specific homologs of subunits of TFIID. Mediator, tMAC and the tTAFs co-regulate expression of a major set of spermatid differentiation genes. The Mediator subunit Med22 binds the tMAC component Topi when the two are coexpressed in S2 cells, suggesting direct recruitment. Loss of Med22 function in spermatocytes causes meiosis I maturation arrest male infertility, similar to loss of function of the tMAC subunits or the tTAFs. Our results illuminate how cell type specific versions of the Mip/dREAM complex and the general transcription machinery cooperate to drive selective gene activation during differentiation in stem cell lineages.


Clustering of discrete cell properties essential for tumorigenicity and metastasis. III. Dissociation of the properties in N-ras-transfected RSV-SR-transformed cells.
We have previously shown that RSV-SR-transformed hamster cells acquire high resistance to H2O2, i.e. the cytotoxic product of activated macrophages (H2O2R) and that they begin to secrete PGE (PGES), thus inactivating the CTA of NK cells. Among normal cells, the same phenotype is expressed in activated macrophages. In all our RSV-transformed cells these 2 properties were jointly expressed and correlated with high tumorigenicity and experimental metastasizing of these cells. We now show that transfection of 3 RSV-SR-transformed cell strains with activated N-ras leads either to complete inhibition of the H2O2R + PGES phenotype in all clones of one strain, or to inhibition of PGES only in the majority of clones of 2 other strains. Unexpectedly, the complete or partial inhibition of this phenotype did not alter the high tumorigenicity of 2 strains of these cells, but lower tumorigenicity was evident in almost all clones of the third strain (as well as in some gene-neo-transfected clones of these strains). The loss of PGES made these cells susceptible to the CTA of NK cells, while the loss of H2O2R did not alter their resistance to the CTA of macrophages. Expression of the H2O2R + PGES phenotype was retained in all cloned variants of control, gene-neo-transfected cells. The possible relation of the N-ras gene to regulation of src gene activities in RSV-SR-transformed cells is discussed.


Skeletal muscle CK-B activity in neurogenic muscular atrophies.
Creatine kinase isoenzymes were determined in skeletal muscle biopsy specimens of 34 patients suffering from neurogenic muscular atrophies. The findings were compared: (1) with those of 38 control muscle samples and (2) with those in 41 muscular dystrophies and other myopathic conditions. The measurements were made by electrophoretic separation and elution of the isoenzymes and by immunoinhibition assay. The results showed that the total and specific CK activity were significantly decreased (P less than 0.005) in neurogenic atrophies in contrast to myopathic conditions where no differences from control levels were observed. This decrease was due to a decrease of the CK-M subunit activity, while the CK-B subunit was elevated. The muscle CK-MB activity was considerably elevated in muscular dystrophies (P less than 0.02) and myositis (P less than 0.001), but it was also slightly elevated in neurogenic conditions. The similarity of the muscle CK isoenzyme pattern in neurogenic atrophies and myotonic dystrophy was noted. These findings could possibly reflect considerable difference in the regeneration process of neurogenic atrophies and muscular dystrophies.


Light and electron microscopic histochemistry of the monoamines in the human foetal sympathetic ganglion in culture.
Sympathetic ganglia of 13 to 19-week-old human foetuses were cultured in small pieces with and without nerve growth factor for up to 5 weeks in vitro. The cultures were studied using phase-contrast, fluorescence and electron microscopy. Monoamines were demonstrated with the formaldehyde-induced fluorescence method, with and without pretreatment of the cultures with catecholamines or monoamine oxidase inhibitor. In the long-term cultures, primitive sympathetic cells, sympathicoblasts of types I and II, and young sympathetic neurons showed a fine structure identical to that described earlier in vivo. There were virtually no satellite or Schwann cells in the cultures. The neurons showed a considerable capacity to grow new nerve fibres in culture, even without nerve growth factor. Nerve terminals with accumulations of synaptic vesicles were regularly observed, occasionally in synapse-like contact with other nervous structures. Large granular vesicles were regularly found in the sympathicoblasts after glutaraldehyde-osmium tetroxide fixation. After permanganate fixation, dense-cored vesicles typical of adrenergic neurons were not seen, either in the perikarya, or in the processes, although it was possible to demonstrate specific fluorescence. No small intensity fluorescent (SIF) cells were observed. Variable formaldehyde-induced fluorescence was observed in the nerve cell perikarya and nerve fibres. The intensity of the fluorescence increased after treatment of the cultures with monoamine oxidase inhibitor and after incubation with catecholamines.


Sequential Liver, Kidney, and Autologous Stem Cell Transplant for AL Amyloidosis: A Case Report.
AL type amyloidosis is a systemic disease characterized by the accumulation of amyloid fibrils that can affect many organs such as the skin, gastrointestinal tract, heart, lungs, liver, and kidney. The most frequently involved organ in amyloidosis is the kidney, but cardiac amyloidosis with the poor prognosis is amyloid organ involvement. In this study, we present the treatment of a 40-year-old female patient with acute Budd-Chiari syndrome and very severe proteinuria with sequential liver, kidney, and autologous stem cell transplant after the diagnosis of systemic amyloidosis. To reduce the effects of massive proteinuria and very severe hypoalbuminemia, bilateral renal artery embolization was performed first. After the evaluation of the patient, she underwent liver transplant from a deceased donor, and then kidney transplant was performed from her son 1 month later. Afterward, the patient was discharged without any problems and underwent chemotherapy and stem cell transplant for primary AL amyloidosis. She was followed up without any problem in terms of liver, kidney, and stem cell at the 24th postoperative month. This case shows that autologous stem cell transplant after kidney and liver transplant may be a good treatment option in a selected patient with stem cell involvement diagnosed as having AL amyloidosis.


Utility of video-assisted method for identifying and preserving the external branch of the superior laryngeal nerve during thyroidectomy.
The EBSLN is vulnerable to damage during thyroidectomy, results in voice related complications, negatively affect patient quality of life, routine identification of the EBSLN prior to surgical manipulation is necessary for a complication-free thyroidectomy. We aimed to validate a video-assisted procedure for identifying and preserving the external branch of the superior laryngeal nerve (EBSLN) during thyroidectomy and analyze the EBSLN Cernea classification and the localization of the nerve entry point (NEP) from the insertion of the sternothyroid muscle. A prospective descriptive study was performed; 134 patients, who scheduled for lobectomy with an intraglandular tumor max diameter ≤ 4 cm and without extrathyroidal extension, were randomly divided into the video-assisted surgery (VAS) and conventional open surgery (COS) groups. We used the video-assisted surgical procedure for visually identifying the EBSLN directly, and compared the differences in the visual identification rate and total identification rate of the two groups. We also measured the localization of the NEP using the insertion of the sternothyroid muscle as a reference. There was no statistically significant difference in clinical characteristics between the two groups. The visual identification rate and total identification rate were significantly higher in the VAS group than the COS group (91.04% vs. 77.61%, 100% vs. 89.6%). The EBSLN injury rate was 0 in both groups. The mean vertical distance (VD) of the NEP from the sternal thyroid insertion was 1.18 mm (SD 1.12 mm, range, 0-5 mm), with approximately 88.97% of the results occurring within the 0-2 mm range. The mean horizontal distance (HD) was 9.33 mm (SD 5.03 mm, range, 0-30 mm), with over 92.13% of the results occurring within the 5-15 mm range. Both the visual and total identification rates of the EBSLN were significantly higher in the VAS group. This method provided a good visual exposure rate of the EBSLN, aiding in identifying and protecting the EBSLN during thyroidectomy.


Hypoxic retinal Muller cells promote vascular permeability by HIF-1-dependent up-regulation of angiopoietin-like 4.
Vision loss from ischemic retinopathies commonly results from the accumulation of fluid in the inner retina [macular edema (ME)]. Although the precise events that lead to the development of ME remain under debate, growing evidence supports a role for an ischemia-induced hyperpermeability state regulated, in part, by VEGF. Monthly treatment with anti-VEGF therapies is effective for the treatment of ME but results in a major improvement in vision in a minority of patients, underscoring the need to identify additional therapeutic targets. Using the oxygen-induced retinopathy mouse model for ischemic retinopathy, we provide evidence showing that hypoxic Müller cells promote vascular permeability by stabilizing hypoxia-inducible factor-1α (HIF-1α) and secreting angiogenic cytokines. Blocking HIF-1α translation with digoxin inhibits the promotion of endothelial cell permeability in vitro and retinal edema in vivo. Interestingly, Müller cells require HIF--but not VEGF--to promote vascular permeability, suggesting that other HIF-dependent factors may contribute to the development of ME. Using gene expression analysis, we identify angiopoietin-like 4 (ANGPTL4) as a cytokine up-regulated by HIF-1 in hypoxic Müller cells in vitro and the ischemic inner retina in vivo. ANGPTL4 is critical and sufficient to promote vessel permeability by hypoxic Müller cells. Immunohistochemical analysis of retinal tissue from patients with diabetic eye disease shows that HIF-1α and ANGPTL4 localize to ischemic Müller cells. Our results suggest that ANGPTL4 may play an important role in promoting vessel permeability in ischemic retinopathies and could be an important target for the treatment of ME.


Differential expression of individual genes encoding the small subunit of ribulose-1,5-bisphosphate carboxylase in Lemna gibba.
The gene family encoding the small subunit (SSU) of ribulose-1,5-bisphosphate carboxylase/oxygenase in the monocot Lemna gibba contains approximately twelve members. We have isolated six of these genes from a genomic library, and sequenced five of the coding regions. The transit peptide nucleotide sequences are conserved, but less highly than the mature polypeptide coding sequence. The mature polypeptide amino acid sequences are identical to each other and to the sequence deduced from a cDNA clone derived from a seventh gene. Each of the five fully characterized genomic sequences contains a single intron in precisely the same position as the second intron of several dicots. The intron sequences differ in length and are less conserved than the coding sequences. The 3'-untranslated regions of the different genes have been sequenced and used to prepare gene-specific probes. These probes have been used to study the expression levels of individual rbcS sequences. Expression of six of the seven genes can be detected in total RNA isolated from plants grown in continuous light. The levels of RNA encoded by each expressed gene are regulated by the action of phytochrome, but there is variability in the amount of expression of each RNA.


Local immunotherapy with the RNA-based immune stimulator CV8102 induces substantial anti-tumor responses and enhances checkpoint inhibitor activity.
Immunotherapy has revolutionized cancer treatment in recent years. Although currently approved checkpoint inhibitors (CPIs) yield remarkable anti-tumoral responses in several cancer types, a substantial proportion of patients do not benefit from such therapies. Local activation of innate immune signaling pathways is a promising approach to overcome the immunosuppressive tumor microenvironment, induce anti-tumor immunity, and improve the efficacy of CPI therapies. Here, we assessed the mode of action and efficacy of the RNA-based innate immune stimulator CV8102 for local immunotherapy in preclinical models. Intratumoral (i.t.) administration of CV8102 activated innate immune responses in the tumor microenvironment and draining lymph nodes, resulting in a dose-dependent anti-tumoral response. Combining i.t. CV8102 with systemic anti-programmed death protein 1 (PD-1) treatment further enhanced anti-tumoral responses, inducing tumor infiltration and activation of CD8+ T cells. The resulting memory response prevented tumor growth in rechallenged animals and impaired the growth of non-injected distal tumors. Therefore, i.t. CV8102 delivery is a promising approach for local cancer immunotherapy, especially in combination with CPIs. Clinical testing of CV8102 is ongoing (NCT03291002).


Concept model of the formation process of humic acid-kaolin complexes deduced by trichloroethylene sorption experiments and various characterizations.
To explore the interactions between soil organic matter and minerals, humic acid (HA, as organic matter), kaolin (as a mineral component) and Ca(2+) (as metal ions) were used to prepare HA-kaolin and Ca-HA-kaolin complexes. These complexes were used in trichloroethylene (TCE) sorption experiments and various characterizations. Interactions between HA and kaolin during the formation of their complexes were confirmed by the obvious differences between the Qe (experimental sorbed TCE) and Qe_p (predicted sorbed TCE) values of all detected samples. The partition coefficient kd obtained for the different samples indicated that both the organic content (fom) and Ca(2+) could significantly impact the interactions. Based on experimental results and various characterizations, a concept model was developed. In the absence of Ca(2+), HA molecules first patched onto charged sites of kaolin surfaces, filling the pores. Subsequently, as the HA content increased and the first HA layer reached saturation, an outer layer of HA began to form, compressing the inner HA layer. As HA loading continued, the second layer reached saturation, such that an outer-third layer began to form, compressing the inner layers. In the presence of Ca(2+), which not only can promote kaolin self-aggregation but can also boost HA attachment to kaolin, HA molecules were first surrounded by kaolin. Subsequently, first and second layers formed (with inner layer compression) via the same process as described above in the absence of Ca(2+), except that the second layer continued to load rather than reach saturation, within the investigated conditions, because of enhanced HA aggregation caused by Ca(2+).


Genetic assessment of age-associated Alzheimer disease risk: Development and validation of a polygenic hazard score.
Identifying individuals at risk for developing Alzheimer disease (AD) is of utmost importance. Although genetic studies have identified AD-associated SNPs in APOE and other genes, genetic information has not been integrated into an epidemiological framework for risk prediction. Using genotype data from 17,008 AD cases and 37,154 controls from the International Genomics of Alzheimer's Project (IGAP Stage 1), we identified AD-associated SNPs (at p < 10-5). We then integrated these AD-associated SNPs into a Cox proportional hazard model using genotype data from a subset of 6,409 AD patients and 9,386 older controls from Phase 1 of the Alzheimer's Disease Genetics Consortium (ADGC), providing a polygenic hazard score (PHS) for each participant. By combining population-based incidence rates and the genotype-derived PHS for each individual, we derived estimates of instantaneous risk for developing AD, based on genotype and age, and tested replication in multiple independent cohorts (ADGC Phase 2, National Institute on Aging Alzheimer's Disease Center [NIA ADC], and Alzheimer's Disease Neuroimaging Initiative [ADNI], total n = 20,680). Within the ADGC Phase 1 cohort, individuals in the highest PHS quartile developed AD at a considerably lower age and had the highest yearly AD incidence rate. Among APOE ε3/3 individuals, the PHS modified expected age of AD onset by more than 10 y between the lowest and highest deciles (hazard ratio 3.34, 95% CI 2.62-4.24, p = 1.0 × 10-22). In independent cohorts, the PHS strongly predicted empirical age of AD onset (ADGC Phase 2, r = 0.90, p = 1.1 × 10-26) and longitudinal progression from normal aging to AD (NIA ADC, Cochran-Armitage trend test, p = 1.5 × 10-10), and was associated with neuropathology (NIA ADC, Braak stage of neurofibrillary tangles, p = 3.9 × 10-6, and Consortium to Establish a Registry for Alzheimer's Disease score for neuritic plaques, p = 6.8 × 10-6) and in vivo markers of AD neurodegeneration (ADNI, volume loss within the entorhinal cortex, p = 6.3 × 10-6, and hippocampus, p = 7.9 × 10-5). Additional prospective validation of these results in non-US, non-white, and prospective community-based cohorts is necessary before clinical use. We have developed a PHS for quantifying individual differences in age-specific genetic risk for AD. Within the cohorts studied here, polygenic architecture plays an important role in modifying AD risk beyond APOE. With thorough validation, quantification of inherited genetic variation may prove useful for stratifying AD risk and as an enrichment strategy in therapeutic trials.


Hippocampal activation and incorporation of macromolecule precursors.
The effect of rhythmic slow wave activity (theta rhythm) on the incorporation of 3H-leucine and 3H-fucose into the total proteins of different hippocampus areas was studied. The theta rhythm was elicited by electrical stimulation of medial septum nuclei. An increase in 3H-leucine incorporation into the total proteins of CA 3 and CA 1 sectors of the hippocampus was observed, whereas the stimulation had no influence on precursor incorporation into the complex CA 4/area dentata. In contrast to these findings 3H-fucose incorporation into hippocampal proteins was not influenced by electrical stimulation of the medial septum. These findings are discussed in comparison to the results obtained in a learning experiment, which revealed an increased incorporation of both leucine and focuse into hippocampal proteins.


Antigenic Fingerprinting of Respiratory Syncytial Virus (RSV)-A-Infected Hematopoietic Cell Transplant Recipients Reveals Importance of Mucosal Anti-RSV G Antibodies in Control of RSV Infection in Humans.
Respiratory syncytial virus (RSV) infection causes significant morbidity in hematopoietic cell transplant (HCT) recipients. However, antibody responses that correlate with recovery from RSV disease are not fully understood. In this study, antibody repertoire in paired serum and nasal wash samples from acutely RSV-A-infected HCT recipients who recovered early (<14 days of RSV shedding) were compared with late-recovered patients (≥14 days of shedding) using gene fragment phage display libraries and surface plasmon resonance. Anti-F serum responses were similar between these 2 groups for antibody repertoires, neutralization titers, anti-F binding antibodies (prefusion and postfusion proteins), antibody avidity, and binding to specific antigenic sites. In contrast, nasal washes from early-recovered individuals demonstrated higher binding to F peptide containing p27. While the serum RSV G antibody repertoires in the 2 groups were similar, the strongest difference between early-recovered and late-recovered patients was observed in the titers of nasal wash antibodies, especially binding to the central conserved domain. Most importantly, a significantly higher antibody affinity to RSV G was observed in nasal washes from early-recovered individuals compared with late-recovered HCT recipients. These findings highlight the importance of mucosal antibodies in resolution of RSV-A infection in the upper respiratory tract.


Generation of effector cells from T cell subsets. I. Similar requirements for Lyt T cell subpopulations in the generation of alloreactive and H-2 restricted killer cells.
Lyt T cell subsets involved in the generation of H-2 restricted and alloreactive cytotoxic effector cells were analysed using anti Lyt antisera. Our data show that Lyt 1, 2, 3+ T cells are required for the induction of primary and secondary H-2 restricted and TNP-specific killer cells. In contrast, primary and secondary H-2 restricted and virus-specific T effector cells were obtained from selected Lyt 2, 3+ T cell populations and were not dependent on the presence of Lyt 1, 2, 3+ T cells. Allogeneic responses to selected K, I, or D region differences were obtained only in the presence of Lyt 1, 2, 3+ T cells; yet alloreactive killer cells were effectively generated from selected Lyt 2, 3+ T cell popluations deprived of Lyt 1, 2, 3+ T cells, when responder and stimulator cells differed at either K + D, K + I, I and D regionre is no qualitative difference between alloreactive and H-2 restricted cytotoxic responses in their requirements for particular Lyt T cell subsets. The findings indicate that the number of different antigenic determinants rather than their association with MHC self determinants is critical for the requirement of Lyt 1, 2, 3+ T cells during the sensitization phase.


Scanning tunneling microscopy and spectroscopy of rubrene on clean and graphene-covered metal surfaces.
Rubrene (C42H28) was adsorbed with submonolayer coverage on Pt(111), Au(111), and graphene-covered Pt(111). Adsorption phases and vibronic properties of C42H28 consistently reflect the progressive reduction of the molecule-substrate hybridization. Separate C42H28 clusters are observed on Pt(111) as well as broad molecular resonances. On Au(111) and graphene-covered Pt(111) compact molecular islands with similar unit cells of the superstructure characterize the adsorption phase. The highest occupied molecular orbital of C42H28 on Au(111) exhibits weak vibronic progression while unoccupied molecular resonances appear with a broad line shape. In contrast, vibronic subbands are present for both frontier orbitals of C42H28 on graphene. They are due to different molecular vibrational quanta with distinct Huang-Rhys factors.


Physicochemical properties of Shiga toxigenic Escherichia coli.
To investigate the physicochemical surface properties, such as cellular surface charge, hydrophobicity and electron donor/acceptor potential of a selection of Shiga toxigenic Escherichia coli (STEC) isolates grown in broth and agar culture. Cellular surface charge was determined using zeta potential measurements. Hydrophobicity of the isolates was determined using bacterial adhesion to hydrocarbons assay, hydrophobic interaction chromatography and contact angle measurements. Microbial adhesion to solvents was used to determine the electron donor/acceptor characteristics. No differences of surface charge measurements were found between broth and agar grown cultures. Isolates belonging to serogroup O157 and serotypes O26:H11 and O111:H- were significantly (P < 0.05) less negatively charged than other STEC serotypes tested. All strains were hydrophilic with most methods and demonstrated a lower hydrophobicity in agar culture compared with broth culture. All strains demonstrated a strong microbial adhesion to chloroform indicating that STEC possess an electron donor and basic character. A relationship between serogroup O157 and other STEC serotypes was apparent using principal-component analysis (PCA). Combining the results for physicochemical properties using PCA differentiated between strains belonging to the O157 serogroup and other STEC/non-STEC strains. PCA found similar results for broth and agar grown cultures. Particular serotypes of STEC possess similar physicochemical properties which may play a role in their pathogenicity or potential attachment to various surfaces.


A new, semi-automated system for the micro-scale synthesis of [195mPt]cisplatin suitable for clinical studies.
A new, semi-automated system for the microscale synthesis of [195mPt]cisplatin has been developed. Radiochemical yields of up to 70% of pure [195mPt]cisplatin can be obtained routinely e.g. 555-629 MBq (15-17 mCi) from 925 MBq (25 mCi) of [195mPt]Pt metal. Chemical losses during synthesis, and radiation exposure of personnel, have been minimized. These increased yields are the consequence of the elimination of most transfers, and enhanced yields at most steps: ammination losses decreased from 27 to 7%, and at the diiodo- to di-acquo- conversion, from 34 to 11%. The versatility of such a system is discussed.


Enzyme-based amperometric galactose biosensors: a review.
This review (with 35 references) summarizes the various strategies used in biosensors for galactose, and their analytical performance. A brief comparison of the enzyme immobilization methods employed and the analytical performance characteristics of a range of galactose biosensors are first summarized in tabular form and then described in detail. Selected examples have been included to demonstrate the various applications of these biosensors to real samples. Following an introduction into the field that covers the significance of sensing galactose in various fields, the review covers biosensors based on the use of galactose oxidase, with a discussion of methods for their immobilization (via cross-linking, adsorption, covalent bonding and entrapment). This is followed by a short section on biosensors based on the use of galactose dehydrogenase. The conclusion section summarizes the state of the art and addresses current challenges. Graphical abstractFabrication of a disposable screen-printed (a) electrochemical galactose biosensor (b) for real sample analysis and a dummy biosensor


A systematic review of compliance to oral nutritional supplements.
Oral nutritional supplements (ONS) play a key role in the management of malnutrition. This systematic review examined patients' compliance with ONS across healthcare settings and the influence of patient and ONS-related factors. A systematic review identified 46 studies (n = 4328) of ONS in which data on compliance (% of prescribed quantity of ONS consumed) was available. Pooled mean %compliance was assessed overall and according to study design and healthcare setting. Inter-relationships between compliance and ONS-related and patient-related factors, and total energy intake were assessed. Overall mean compliance with ONS was 78% (37%-100%; 67% hospital, 81% community; overall mean ONS intake 433 kcal/d). Percentage compliance was similar in randomised (79%) and non-randomised (77%) trials, with little variation between diagnostic groups. Compliance across a heterogeneous group of unmatched studies was positively associated with higher energy-density ONS and greater ONS and total energy intakes, negatively associated with age, and unrelated to amount or duration of ONS prescription. This systematic review suggests that compliance to ONS is good, especially with higher energy-density ONS, resulting in improvements in patients' total energy intakes that have been linked with clinical benefits. Further research is required to address the compliance and effectiveness of other common methods of oral nutritional support.


Design and synthesis of tetrahydrophthalimide derivatives as inhibitors of HIV-1 reverse transcriptase.
Non-nucleoside reverse transcriptase inhibitors (NNRTIs) are one of the key components in highly active anti-retroviral therapy because of their high specificity and less toxicity. NNRTIs inhibit reverse transcriptase enzyme by binding to the allosteric site, which is 10Å away from the active site. Rapid emergence of resistance is the major problem with all anti-HIV agents. Hence, there is continuous need to develop novel anti-HIV agents active against both drug sensitive and resistance strains. All the 16 synthesized 2-(1,3-dioxo-3a,4-dihydro-1H-isoindol-2(3H,7H,7aH)-yl)-N-(substitutedphenyl) acetamide 4(a-p) analogs were characterized by Fourier transform infrared spectroscopy, proton nuclear magnetic resonance spectroscopy, mass spectroscopy, and elemental analysis. Lipinski rule of five parameters and molecular parameters like solubility, drug likeness, and drug score were derived for designed analogs using online servers like Molinspiration and Osiris property explorer. Synthesized compounds were evaluated for their HIV-1 reverse transcriptase inhibitor activity by HIV-1 RNA-dependent DNA polymerase activity assay at 2 and 20 μM concentrations. Among the 16 synthesized compounds, 4a, 4b, 4f, 4g, 4k, and 4l showed weak reverse transcriptase inhibitor activity at 20 μM concentration. For the designed compounds, there was no correlation observed between molecular modeling and in vitro studies.


Minimal astigmatism after sutureless planned extracapsular cataract extraction.
To evaluate astigmatism after mini-nuc extracapsular cataract extraction (ECCE) in which a chevron incision is enlarged to 6.0 to 7.0 mm for easier nucleus removal and to compare the results with those using a 5.0 mm incision. Department of Ophthalmology, The Edith Wolfson Medical Center, Sackler Faculty of Medicine, Tel-Aviv University, Holon, Israel. Thirty eyes of 29 patients were enrolled in this study. Keratometry was performed preoperatively and 3 to 9 months postoperatively. The incision length was 6.0 mm in 6 eyes, 6.5 mm in 10 eyes, and 7.0 mm in 14 eyes that had mature cataract. The mean induced astigmatism calculated by simple subtraction was 0.12 diopter (D) +/- 0.51 (SD), 0.16 +/- 0.98 D, and 0.67 +/- 0.91 D for the 6.0 mm, 6.5 mm, and 7.0 mm incision, respectively. By vector analysis, the mean induced astigmatism was 0.60 +/- 0.30 D, 0.75 +/- 0.67 D, and 1.36 +/- 0.77 D, respectively. Results by both methods showed no significant difference between the previously reported 5.0 mm incision and the 6.0 mm and 6.5 mm incisions. The 7.0 mm group had statistically significantly greater induced astigmatism than the 5.0 mm group (P =.01, simple subtraction; P =.002, vector analysis). Enlarging the size of the chevron incision up to 7.0 mm resulted in a small increase in induced astigmatism. The enlarged incision simplified the operative technique.


Influence of Iron Deficiency on Clinical and Haemodynamic Parameters in Pulmonary Arterial Hypertension Cohorts.
Iron deficiency (Fedef) has been shown to be common in patients with group 1 or pulmonary arterial hypertension (PAH). Several studies have shown a negative impact of Fedef on clinical and haemodynamic parameters of the disease, but data from individual studies have not been strong enough to lead to incorporation of the finding of Fedef into prognostic or therapeutic algorithms. The goal of this meta-analysis was to combine data from available studies to better define any associations between Fedef and established variables of prognostic importance in PAH. A literature search identified nine studies with extractable data relevant to the study questions. The impact of Fedef upon the following parameters was evaluated: 6-minute walk distance (6MWD), WHO-functional class, N-terminal pro-brain natriuretic peptide (NT-proBNP) levels, echocardiography, and findings from right heart catheterisation (RHC). Pooled results were reported as mean difference or risk difference with 95% confidence intervals utilising a random effects modeling approach. Fedef in the PAH population was common (47% of cases) and was associated with cardiovascular dysfunction (lower tricuspid annular plane systolic excursion [TAPSE], elevated NT-proBNP, and lower mixed venous oxygen saturation) and with reduction in functional capacity (lower 6MWD and higher functional class). This meta-analysis strengthens the relationships between Fedef and several markers of poor outcome in PAH. Fedef in patients with PAH warrants further scrutiny and merits consideration as a cause of clinical deterioration. Even though causation and longitudinal relationships between Fedef and PAH could not be identified, effect of Fedef on factors that affect disease prognosis is noteworthy and worthy of more focussed studies.


The mental health of Muslim mothers in extended families living in Britain: the impact of intergenerational disagreement on anxiety and depression.
The study assessed the impact of intergenerational differences of opinion over child rearing on the mental health of Muslim mothers living in extended families. The study adopted a correlational design in an attempt to identify factors that accounted for mental health problems. The child-rearing attitudes of mothers and grandmothers, mothers' mental health, levels of family acculturation and a range of other background and demographic information was collected from 54 extended families living in two Muslim communities in London using Urdu versions of standard questionnaires. Rates of depression and anxiety among the mothers in the study were high. Grandmothers had more traditional attitudes to child rearing than did mothers. Intergenerational discrepancy over child rearing was more marked in more acculturated families. Discrepancy was associated with higher levels of mothers' anxiety and depression. The unusually high levels of depression and anxiety displayed by Muslim mothers living in extended families can in part be accounted for by patterns of intergenerational discrepancy. These possibly reflect discordant world views within those families that have been assimilated into the dominant British culture.


Switching therapy from intravenous beta blocker to bisoprolol transdermal patch for atrial fibrillation tachycardia.
Beta-blockers are important for severe-status patients with atrial fibrillation-related tachycardia. Beta 1-selective intravenous injection are routinely used, but long-term administration is difficult due to cost-performance- or management-related issues. A bisoprolol patch, a beta-blocker to be percutaneously absorbed, recently became commercially available in Japan. As it may facilitate effective absorption and a mild elevation of the blood concentration, we retrospectively analyzed 16 patients with atrial fibrillation-related tachycardia who were admitted to the Intensive Care Unit of Hitachi General Hospital Emergency and Critical Care Center, and underwent switching therapy from landiolol to a bisoprolol patch. For switching, the bisoprolol patch (4 mg) was attached to each patient. The bisoprolol patch was introduced 88 h after the start of landiolol administration, when the rate was approximately 3 μg/kg/min. Landiolol injection was combined with bisoprolol for 15.4 ± 17.5 h. Switching therapy was successful in all subjects. The introduction of the bisoprolol patch did not induce any significant changes in the blood pressure or heart rate. After the completion of landiolol administration, there were also no significant changes in either parameter. There were no adverse events. In severe-status patients, switching therapy from landiolol injection to the bisoprolol patch can be conducted safely, and might be useful for heart-rate control.


Navafenterol (AZD8871) in healthy volunteers: safety, tolerability and pharmacokinetics of multiple ascending doses of this novel inhaled, long-acting, dual-pharmacology bronchodilator, in two phase I, randomised, single-blind, placebo-controlled studies.
Navafenterol (AZD8871) is a novel, long-acting, dual-pharmacology (muscarinic receptor antagonist and β2-adrenoceptor agonist) molecule in development for chronic obstructive pulmonary disease and asthma. These two phase I, randomised, single-blind, multiple-ascending-dose studies evaluated inhaled navafenterol and placebo (3:1 ratio) in healthy, male, non-Japanese (study A; NCT02814656) and Japanese (study B; NCT03159442) volunteers. In each study, volunteers were dosed in three cohorts, allowing gradual dose escalation from 300 μg to 600 μg to 900 μg. The primary objective was to investigate the safety and tolerability of navafenterol at steady state. Pharmacokinetics were also assessed. Twenty-four volunteers completed each study (navafenterol, n = 6; placebo, n = 2 in each cohort). There were no deaths, serious adverse events (AEs) or treatment-emergent AEs (TEAEs) leading to discontinuation of navafenterol. The most frequent TEAEs were vessel puncture-site bruise (placebo, n = 2; navafenterol 900 μg; n = 3) in study A and diarrhoea (placebo, n = 1; navafenterol 300 μg, n = 2; navafenterol 900 μg, n = 3) in study B. No dose-response relationship was observed for TEAEs. There was a dose-dependent increase in mean heart rate on day 16 in both studies. The pharmacokinetics of navafenterol were similar between non-Japanese and Japanese volunteers. Multiple ascending doses of navafenterol were well-tolerated and the safety and pharmacokinetics of navafenterol were similar in non-Japanese and Japanese volunteers. The findings support navafenterol clinical development. ClinicalTrials.gov ; Nos.: NCT02814656 and NCT03159442; URL: www.clinicaltrials.gov .


Abnormal Bone Acquisition With Early-Life HIV Infection: Role of Immune Activation and Senescent Osteogenic Precursors.
Chronic immune activation associated with human immunodeficiency virus (HIV) infection may have negative consequences on bone acquisition in individuals infected with HIV early in life. Bone mineral density (BMD) and microarchitecture were characterized in 38 HIV-infected men on antiretroviral therapy (18 perinatally-infected, 20 adolescence-infected) and 20 uninfected men age 20 to 25 years by dual-energy X-ray absorptiometry (DXA), high-resolution peripheral quantitative computed tomography (HRpQCT). Flow cytometry was utilized to measure CD4+/CD8+ activation (HLADR+CD38+) and senescence (CD28-CD57+) and to quantify circulating osteogenic precursor (COP) cells in peripheral blood mononuclear cells using antibodies to RUNX2 and osteocalcin (OCN). Telomere lengths were measured in sorted COP cells using qPCR. DXA-derived areal BMD Z-scores and HRpQCT-derived volumetric BMD (vBMD) measures were lower in HIV-infected than uninfected men. Proportion of activated and senescent CD4+ and CD8+ T cells were higher in HIV-infected than uninfected men. The percentage of COP cells (mean ± SE) was lower in HIV-infected than uninfected (0.19% ± 0.02% versus 0.43% ± 0.06%; p < 0.0001) men, and also lower in perinatally-infected than adolescence-infected men (0.15% ± 0.02% versus 0.22% ± 0.03%; p < 0.04). A higher proportion of COP cells correlated with higher bone stiffness, a measure of bone strength, whereas a higher proportion of activated CD4+ T cells correlated with lower BMD and stiffness and lower proportion of COP cells. T cell activation with HIV-infection was associated with decreased numbers of osteogenic precursors as well as lower peak bone mass and bone strength. © 2016 American Society for Bone and Mineral Research.


Juvenile Idiopathic Arthritis: A Review of Novel Diagnostic and Monitoring Technologies.
Juvenile idiopathic arthritis (JIA) is the most common rheumatic disease of childhood and is characterized by an often insidious onset and a chronic relapsing-remitting course, once diagnosed. With successive flares of joint inflammation, joint damage accrues, often associated with pain and functional disability. The progressive nature and potential for chronic damage and disability caused by JIA emphasizes the critical need for a prompt and accurate diagnosis. This article provides a review of recent studies related to diagnosis, monitoring and management of JIA and outlines recent novel tools and techniques (infrared thermal imaging, three-dimensional imaging, accelerometry, artificial neural networks and fuzzy logic) which have demonstrated potential value in assessment and monitoring of JIA. The emergence of novel techniques to assist clinicians' assessments for diagnosis and monitoring of JIA has demonstrated promise; however, further research is required to confirm their clinical utility.


Identification and characterization of the tuberous sclerosis gene on chromosome 16.
Tuberous sclerosis (TSC) is an autosomal dominant multisystem disorder with loci assigned to chromosomes 9 and 16. Using pulsed-field gel electrophoresis (PFGE), we identified five TSC-associated deletions at 16p13.3. These were mapped to a 120 kb region that was cloned in cosmids and from which four genes were isolated. One gene, designated TSC2, was interrupted by all five PFGE deletions, and closer examination revealed several intragenic mutations, including one de novo deletion. In this case, Northern blot analysis identified a shortened transcript, while reduced expression was observed in another TSC family, confirming TSC2 as the chromosome 16 TSC gene. The 5.5 kb TSC2 transcript is widely expressed, and its protein product, tuberin, has a region of homology to the GTPase-activating protein GAP3.


Diffuse noxious inhibitory controls of lumbar spinal neurons involve a supraspinal loop in the cat.
In anesthetized cats, lumbar dorsal horn neurons were excited by brief noxious radiant heating of glabrous hindpaw skin. These nociceptive responses were inhibited by concomitant repetitive electrical stimulation of the ipsilateral deep radial nerve. Noxious heat responses were linearly correlated with skin temperature during heating. The slope of this stimulus-response function was decreased, and the response threshold increased, by deep radial nerve stimulation. Microinjection of lidocaine into the medullary raphe attenuated the inhibition induced by deep radial nerve stimulation. The results indicate that in the cat, 'diffuse noxious inhibitory controls' (DNIC) involve medial medullary regions.


A conceptual framework for navigating the superficial territories of the face: Relevant anatomic points for the dermatologic surgeon.
Recent publications in the dermatologic surgery literature have sparked a re-emergence of interest in anatomy relevant for the Mohs surgeon necessitating a re-visit of under-appreciated concepts, regarding the topography of the face and its visceral contents from a surgically relevant perspective. This paper presents a pre-operative review and a conceptual framework for intra-operative planning for Mohs micrographic surgery and reconstruction. The key concepts presented are based on a series of (1) reviews regarding clinically significant points aimed at improving outcomes for reconstructive surgery, (2) anatomical dissections of fresh frozen cadavers, and (3) surgical experience of the authors. Basic anatomical concepts have been assimilated, surgically evaluated and re-directed toward the dermatologic surgeon in the hope that improved anatomic competence will reduce surgical hesitance.


The pin-striped habit balancing charity and business in Catholic hospitals, 1865-1915.
Nursing literature seldom examines Catholic si. ters as entrepreneurs competing in the hospital marketplac even though they established hospitals throughout the United States in the late 19th and early 20th centuries. This study examines the tension between hospita as businesses and charities by asking how nursing sisters, who took vows of poverty, could build impressive hospital networks. What links did they forge to make hospital building possible? Historical methodology is used which draws upon primary sources in archives of three women's religious congregations. These include nuns' constitutions and letters, hospital chronicles, journals, minutes of meetings, annual reports and census records. Secondary sources include nursing hospital, religious, and economic histories. Catholic sisters' entrepreneurial activities were pragmatic adaptions that reflected their commitment to their nursing and hospital establishments. They raised money and formed alliances not to build personal fortunes but to build religious values. Their entrepreneurship and spirituality worked together. Sister-nurses' viewed their actions as doing what was necessary to their mission. A finacially stable hospital, with it's large patient population, enhanced the social and spiritual importance of nun's work. Yet, the broad-based corporate culture of Catholic hospitals still needs further study.


Dexpramipexole improves bioenergetics and outcome in experimental stroke.
Dexpramipexole, a drug recently tested in patients with amyotrophic lateral sclerosis (ALS,) is able to bind F1Fo ATP synthase and increase mitochondrial ATP production. Here, we have investigated its effects on experimental ischaemic brain injury. The effects of dexpramipexole on bioenergetics, Ca2+ fluxes, electrophysiological functions and death were evaluated in primary neural cultures and hippocampal slices exposed to oxygen-glucose deprivation (OGD). Effects on infarct volumes and neurological functions were also evaluated in mice following proximal or distal middle cerebral artery occlusion (MCAo). Distribution of dexpramipexole within the ischaemic brain was evaluated by means of mass spectrometry imaging. Dexpramipexole increased mitochondrial ATP production in cultured neurons or glia and reduces energy failure, prevents intracellular Ca2+ overload and affords cytoprotection when cultures are exposed to OGD. This compound also counteracted ATP depletion, mitochondrial swelling, anoxic depolarization, loss of synaptic activity and neuronal death in hippocampal slices subjected to OGD. Post-ischaemic treatment with dexpramipexole, at doses consistent with those already used in ALS patients, reduced brain infarct size and ameliorated neuroscore in mice subjected to transient or permanent MCAo. Notably, the concentrations of dexpramipexole reached within the ischaemic penumbra equalled those found neuroprotective in vitro. Dexpramipexole, a compound able to increase mitochondrial F1Fo ATP-synthase activity reduced ischaemic brain injury. These findings, together with the excellent brain penetration and favourable safety profile in humans, make dexpramipexole a drug with realistic translational potential for the treatment of stroke. This article is part of a themed section on Inventing New Therapies Without Reinventing the Wheel: The Power of Drug Repurposing. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.2/issuetoc.


Regional clinical practice patterns in reproductive endocrinology: a collaborative transnational pilot survey of in vitro fertilization programs in the Middle East.
This research describes current clinical and demographic features sampled from reproductive endocrinology programs currently offering in vitro fertilization (IVF) in the Middle East. Clinic leadership provided data via questionnaire on patient demographics, demand for IVF services, annual cycle volume, indications for IVF, number of embryos transferred, twinning frequency, local regulations governing range of available adjunct therapies, time interval between initial enrollment and beginning IVF as well as information about other aspects of IVF at each center. Data were received from representative IVF clinics (n = 13) in Cyprus, Egypt, Iran, Israel, Jordan, Lebanon, Qatar, Saudi Arabia and Turkey. Mean (+/- SD) age of respondents was 47.8 +/- 8 yrs, with average tenure at their facility of 11.2 +/- 6 yrs. Estimated total number of IVF programs in each nation responding ranged from 1 to 91. All respondents reported individual participation in accredited CME activity within 24 months. 76.9% performed embryo transfers personally; blastocyst transfer was available at 84.6% of centers. PGD was offered at all sites. In this population, male factor infertility accounted for most IVF consultations and the majority (59.1%) of female IVF patients were < 35 yrs of age. Prevalence of smoking among female IVF patients was 7.2%. Average number of embryos transferred was 2.4 (+/- 0.4) for patients at age < 35 yrs, and 2.9 (+/- 0.8) at age > 41 yrs. For these age categories, twinning (any type) was observed in 22.6 (+/- 10.8)% and 13.7 (+/- 10.4)%, respectively. In 2005, the average number of IVF cycles completed at study sites was 1194 (range 363-3500) and 1266 (range 263-4000) in 2006. Frozen embryo transfers accounted for 17.2% of cycles at these centers in 2005. Average interval between initial enrollment and IVF cycle start was 8 weeks (range 0.3-3.5 months). This sampling of diverse IVF clinics in the Middle East, believed to be the first of its kind, identified several common factors. Government registry or oversight of clinical IVF practice was limited or nonexistent in most countries, yet number of embryos transferred was nevertheless fairly uniform. Sophisticated reproductive health services in this region are associated with minimal delay (often < 8 weeks) from initial presentation to IVF cycle start. Most Middle East nations do not maintain a comprehensive IVF database, and there is no independent agency to collect transnational data on IVF clinics. Our pilot study demonstrates that IVF programs in the Middle East could contribute voluntarily to collaborative network efforts to share clinical data, improve quality of care, and increase patient access to reproductive services in the region.


Salivary markers in patients with chronic renal failure.
Chronic renal failure (CRF) is a progressive loss of renal function over a period of months or years. The major function of the kidneys is the removal of metabolic waste products, electrolytes and water. When this function is impaired, systemic changes, oral complications and alterations in salivary composition may occur. This study aimed to compare the levels of immunological and inflammatory components in the saliva samples from patients that undergo to hemodialysis treatment (HD), without HD and control. This study evaluated IgA, IgG, C reactive protein (CRP) and nitric oxide (NO) in saliva samples from 119 patients, who were divided into the control group (C), chronic renal failure (CRF) patient group and CRF patients on hemodialysis treatment (HD) group. IgA and IgG levels were analyzed by ELISA. Nitric oxide levels were determined indirectly by the nitrite concentration using Griess reagent; CRP by agglutination tests; and total proteins, by Bradford assay. The HD group showed significantly higher levels of IgG, IgA and CRP compared with the control and CRF groups. The CRF group presented the same amounts of IgG, IgA and CRP as the C group but significantly higher levels of NO similar to the HD group. Renal disease, particularly hemodialysis treatment during renal disease, seems to alter salivary immunological and inflammatory components. Thus, analyzing the levels of IgA, IgG, NO and CRP in saliva may be beneficial for monitoring renal disease.


Limited mandibular movement: causes and treatment.
Limited mandibular movement is a challenging clinical problem. A variety of treatment modalities have been described to permit planning of an individual course of treatment for each patient.


Isolation, Culture, and Transfection of Melanocytes.
Located in the basal epidermis and hair follicles, melanocytes of the integument are responsible for its coloration through production of melanin pigments. Melanin is produced in a type of lysosome-related-organelle (LRO) called the melanosome. In humans, this skin pigmentation acts as an ultraviolet radiation filter. Abnormalities in the division of melanocytes are quite common, with potentially oncogenic growth usually followed by cell senescence producing benign naevi (moles), or occasionally, melanoma. Therefore, melanocytes are a useful model for studying both cellular senescence and melanoma, as well as many other aspects of biology such as pigmentation, organelle biogenesis and transport, and the diseases affecting these mechanisms. Melanocytes for use in basic research can be obtained from a range of sources, including surplus postoperative skin or from congenic murine skin. Here we describe methods to isolate and culture melanocytes from both human and murine skin (including the preparation of mitotically inactive keratinocytes for use as feeder cells). We also describe a high-throughput transfection protocol for human melanocytes and melanoma cells. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Primary explantation of human melanocytic cells Basic Protocol 2: Preparation of keratinocyte feeder cells for use in the primary culture of mouse melanocytes Basic Protocol 3: Primary culture of melanocytes from mouse skin Basic Protocol 4: Transfection of human melanocytes and melanoma cells.


Translating Research into Child Health Policy: Aligning Incentives and Building a New Discourse.
Research has led to major achievements in public policy and child health. Despite the gains, the need for research to inform policy remains paramount against a backdrop of inadequate public health investments, health inequities, and public skepticism toward science. However, the translation of research into child health policy has often been slow due to misalignments in incentives between researchers and policy makers and a paucity of conceptual models to inform translation. This article outlines barriers to translation, provides examples of discordance between evidence and policy, summarizes models to inform translation, and offers strategies to improve translation of research to policy.


Baohuoside i suppresses breast cancer metastasis by downregulating the tumor-associated macrophages/C-X-C motif chemokine ligand 1 pathway.
Breast cancer is the most common malignancy in women and metastasis is the leading cause of breast cancer-related deaths. Our previous studies have shown that XIAOPI formula, a newly approved drug by the State Food and Drug Administration of China (SFDA), can dramatically inhibit breast cancer metastasis by modulating the tumor-associated macrophages/C-X-C motif chemokine ligand 1 (TAMs/CXCL1) pathway. However, the bioactive compound accounting for the anti-metastatic effect of XIAOPI formula remains unclear. This study was designed to separate the anti-metastatic bioactive compound from XIAOPI formula and to elucidate its action mechanisms. TAMs/CXCL1 promoter activity-guided fractionation and multiple chemical structure identification approaches were conducted to screen the bioactive compound from XIAOPI formula. Breast cancer cells and TAMs were co-cultured in vitro or co-injected in vivo to simulate their coexistence. Multiple molecular biology experiments, zebrafish breast cancer xenotransplantation model and mouse breast cancer xenografts were applied to validate the anti-metastatic activity of the screened compound. Bioactivity-guided fractionation identified baohuoside I (BHS) as the key bioactive compound of XIAOPI formula in inhibiting TAMs/CXCL1 promoter activity. Functional studies revealed that BHS could significantly inhibit the migration and invasion as well as the expression of metastasis-related proteins in both human and mouse breast cancer cells, along with decreasing the proportion of breast cancer stem cells (CSCs). Furthermore, BHS could suppress the M2 phenotype polarization of TAMs and therefore attenuate their CXCL1 expression and secretion. Notably, mechanistic investigations validated TAMs/CXCL1 as the crucial target of BHS in suppressing breast cancer metastasis as exogenous addition of CXCL1 significantly abrogated the anti-metastatic effect of BHS on breast cancer cells. Moreover, BHS was highly safe in vivo as it exhibited no observable embryotoxicity or teratogenic effect on zebrafish embryos. More importantly, BHS remarkably suppressed breast cancer metastasis and TAMs/CXCL1 activity in both zebrafish breast cancer xenotransplantation model and mouse breast cancer xenografts. This study not only provides novel insights into TAMs/CXCL1 as a reliable screening target for anti-metastatic drug discovery, but also suggests BHS as a promising candidate drug for metastatic breast cancer treatment.


Additional superdrainage reduces anastomotic fistula and stenosis after gastric tube reconstruction with cervical anastomosis for esophageal cancer.
Gastric pull-up is a common procedure to reconstruct the continuity of the upper digestive tract after esophagectomy. However, this technique sometimes causes postoperative anastomotic leakage or stricture, resulting from congestion of the gastric tube. We performed additional microvascular venous anastomoses to solve this problem. The purpose of this study was to compare postoperative anastomotic leaks and strictures in cases with or without additional venous superdrainage after gastric tube reconstruction. A total of 117 consecutive patients with cervical and thoracic esophageal cancer who underwent thoracoscopic esophagectomy with gastric tube reconstruction in the National Nagasaki Medical Center between 2011 and 2021 were analyzed retrospectively. Of these patients, 46 did not undergo additional venous anastomoses (standard group), and 71 who underwent gastric pull-up surgery after November 2014 have added this surgical procedure to their routine (superdrainage group). We compared the frequency of postsurgical leakage and stricture in the 2 groups retrospectively. Fifteen patients (32.6%) developed postoperative leakage in the standard group and 6 (8.5%) did so in the superdrainage group. Twelve patients (26.1%) showed postoperative anastomotic stricture in the standard group and 7 (9.9%) did so in the superdrainage group. Patients who did not undergo additional venous superdrainage were significantly more likely to develop postsurgical leakage (χ2 test P < .01) and anastomotic stricture (χ2 test P < .05). The mean time taken to perform additional venous anastomoses was 54.2 minutes. Our study revealed that performing additional venous anastomosis for as little as 1 hour can significantly reduce the incidence of postoperative leakage and stenosis. This procedure is of merit to perform after total esophagectomy with gastric tube reconstruction.


Prognostic Risk Stratification and End-of-Life Care Outcomes in Patients With Metastatic Melanoma Treated With Immune Checkpoint Inhibitors.
The emergence of immune checkpoint inhibitors (ICIs) has improved survival outcomes in patients with metastatic melanoma, while potentially increasing the use of systemic therapy near the end of life (EOL). Yet, less is known on how to facilitate treatment decision making and identify patients who might benefit from early palliative care comanagement. We determined baseline clinical and laboratory factors that are associated with poor prognosis for patients with advanced melanoma treated with ICIs. We subsequently identified prognostic subgroups to evaluate association with EOL outcomes and determine if EOL care varied across prognostic strata. Our cohort included 398 patients with metastatic melanoma treated with ICIs. Factors associated with overall survival (OS) included: lactate dehydrogenase, neutrophil/lymphocyte ratio, performance status, prior therapies, liver metastases, and lung metastases. Patients were stratified by risk of death using risk scores developed from multivariable analyses. A total of 205 patients died: 45/133 (34%) low-risk, 63/133 (47%) medium-risk, and 97/132 (73%) of high-risk patients. Among those who died, higher risk patients were more likely to receive ICIs within 14, 30, and 90 days of death. We found no association between risk group and hospice referrals or location of death. Patients with metastatic melanoma at highest risk of death as defined by our model were more likely than lower-risk patients to receive ICIs near the EOL. Prognostic risk stratification may guide early palliative care interventions to appropriately utilize ICIs and optimize EOL care.