Genetic Burden for Late-Life Neurodegenerative Disease and Its Association With Early-Life Lipids, Brain, Behavior, and Cognition.
Genetics play a significant role in the etiology of late-life neurodegenerative diseases like Alzheimer's disease, Parkinson's disease, and frontotemporal dementia. Part of the individual differences in risk for these diseases can be traced back decades before the onset of disease symptoms. Previous studies have shown evidence for plausible links of apolipoprotein E (APOE), the most important genetic marker for Alzheimer's disease, with early-life cognition and neuroimaging markers. We aimed to assess whether genome-wide genetic burden for the aforementioned neurodegenerative diseases plays a role in early-life processes. We studied children from the Generation R Study, a prospective birth cohort. APOE genotypes and polygenic genetic burdens for Alzheimer's disease, Parkinson's disease, and frontotemporal dementia were obtained through genome-wide genotyping. Non-verbal intelligence was assessed through cognitive tests at the research center around the age of 6 years, and educational attainment through a national school performance test around the age of 11 years. The Child Behavior Checklist was administered around the age of 10 years, and data from the anxious/depressed, withdrawn/depressed, and the internalizing behavior problems scales were used. Children participated in a neuroimaging study when they were 10 years old, in which structural brain metrics were obtained. Lipid serum profiles, which may be influenced by APOE genotype, were assessed from venal blood obtained around the age of 6 years. The sample size per analysis varied between 1,641 and 3,650 children due to completeness of data. We did not find evidence that APOE genotype or the polygenic scores impact on childhood nonverbal intelligence, educational attainment, internalizing behavior, and global brain structural measures including total brain volume and whole brain fractional anisotropy (all p > 0.05). Carriership of the APOE ε2 allele was associated with lower and APOE ε4 with higher low-density lipoprotein cholesterol concentrations when compared to APOE ε3/ε3 carriers. We found no evidence that genetic burden for late-life neurodegenerative diseases associates with early-life cognition, internalizing behavior, or global brain structure.


The Sapir-Whorf hypothesis and inference under uncertainty.
The Sapir-Whorf hypothesis holds that human thought is shaped by language, leading speakers of different languages to think differently. This hypothesis has sparked both enthusiasm and controversy, but despite its prominence it has only occasionally been addressed in computational terms. Recent developments support a view of the Sapir-Whorf hypothesis in terms of probabilistic inference. This view may resolve some of the controversy surrounding the Sapir-Whorf hypothesis, and may help to normalize the hypothesis by linking it to established principles that also explain other phenomena. On this view, effects of language on nonlinguistic cognition or perception reflect standard principles of inference under uncertainty. WIREs Cogn Sci 2017, 8:e1440. doi: 10.1002/wcs.1440 For further resources related to this article, please visit the WIREs website.


Effects of passive smoking on outcome in pregnancy.
Adverse health effects of exposure to environmental tobacco smoke (ETS) among non-smokers have been studied occasionally in developing countries. To study the effects of exposure to ETS on outcome in pregnancy A cross-sectional study at a secondary level teaching hospital Material and Methods: Consecutive 576 non-smoking women delivering a singleton live baby were studied. A pre-designed structured questionnaire was used to record the details of exposure to ETS at home. The maternal and foetal variables were compared among those who were exposed to ETS vis-à-vis not exposed. Unpaired Student t-test was used for the comparison of continuous variables and Fisher's Exact test was used for categorical variables. Multiple logistic regression analysis was performed after including all variables found to have significant differences on univariate analysis. Of the 576 women studied 141 (24%) were exposed to ETS. In the mothers exposed to ETS, there was a significantly higher incidence of pre-term birth (24.1% vs. 16.1%; P = 0.027) and small-for-gestation babies (31.9% vs.17.2%; p<0.001) as compared to unexposed mothers. The mean birth weight of the babies born to the mothers exposed to ETS was 138 g less than that of babies in the unexposed group (2632 -/+577 g vs. 2770 -/+562 g respectively, p = 0.014). The multiple logistic regression analyses showed that ETS exposure during pregnancy was significantly associated with a higher risk of small-for-gestation babies (OR 2.10; 95% CI: 1.27-3.48). Exposure to ETS during pregnancy is associated with higher risk of having a small-for-gestation baby.


Economic instability and household chaos relate to cortisol for children in poverty.
The present study investigated relations among various aspects of economic adversity and cortisol levels for young children facing economic hardship. Specifically, the study examined relations to cortisol for variables representing family income, material hardship, financial strain, economic instability, and household chaos. Participants were 374 children, ages 3-5 years, who attended a Head Start preschool, as well as their primary caregivers. Nearly all children lived in households classified as poor or low-income, defined as less than two times the federal poverty threshold. Caregivers completed interviews about family demographics and economic adversity at the beginning of the school year. Child salivary cortisol was sampled in duplicate on two weekday mornings at the end of the school year. We hypothesized that economic instability would show direct statistical effects on child cortisol as well as indirect effects via household chaos. A structural equation model that corresponded to this hypothesis showed adequate fit for the sample data and revealed a statistically significant indirect effect of economic adversity on child cortisol via economic instability and household chaos, as well as statistically significant direct effects of economic instability and chaos on child cortisol, and a significant indirect effect of economic instability on cortisol via household chaos. Implications concern understanding mechanisms of poverty risk, including the impact of instability and chaos on stress physiology, and promoting physiological regulation for children facing economic hardship. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Effects of high medium pH on growth, metabolism and transport in Saccharomyces cerevisiae.
Growth of Saccharomyces cerevisiae stopped by maintaining the pH of the medium in a pH-stat at pH 8.0 or 9.0. Studying its main physiological capacities and comparing cells after incubation at pH 6.0 vs. 8.0 or 9.0, we found that (a) fermentation was moderately decreased by high pH and respiration was similar and sensitive to the addition of an uncoupler, (b) ATP and glucose-6-phosphate levels upon glucose addition increased to similar levels and (c) proton pumping and K(+) transport were also not affected; all this indicating that energy mechanisms were preserved. Growth inhibition at high pH was also not due to a significant lower amino acid transport by the cells or incorporation into proteins. The cell cycle stopped at pH 9.0, probably due to an arrest as a result of adjustments needed by the cells to contend with the changes under these conditions, and microarray experiments showed some relevant changes to this response.


Nitric oxide-peroxynitrite-poly(ADP-ribose) polymerase pathway in the skin.
In the last decade it has become well established that in the skin, nitric oxide (NO), a diffusable gas, mediates various physiologic functions ranging from the regulation of cutaneous blood flow to melanogenesis. If produced in excess, NO combines with superoxide anion to form peroxynitrite (ONOO-), a cytotoxic oxidant that has been made responsible for tissue injury during shock, inflammation and ischemia-reperfusion. The opposite effects of NO and ONOO- on various cellular processes may explain the 'double-edged sword' nature of NO depending on whether or not cellular conditions favour peroxynitrite formation. Peroxynitrite has been shown to activate the nuclear nick sensor enzyme, poly(ADP-ribose) polymerase (PARP). Overactivation of PARP depletes the cellular stores of NAD+, the substrate of PARP, and the ensuing 'cellular energetic catastrophy' results in necrotic cell death. Whereas the role of NO in numerous skin diseases including wound healing, burn injury, psoriasis, irritant and allergic contact dermatitis, ultraviolet (UV) light-induced sunburn erythema and the control of skin infections has been extensively documented, the intracutaneous role of peroxynitrite and PARP has not been fully explored. We have recently demonstrated peroxynitrite production, DNA breakage and PARP activation in a murine model of contact hypersensitivity, and propose that the peroxynitrite-PARP route represents a common pathway in the pathomechanism of inflammatory skin diseases. Here we briefly review the role of NO in skin pathology and focus on the possible roles played by peroxynitrite and PARP in various skin diseases.


[Spin label study of slow molecular rotations of globular proteins using microwave saturation effects in electron paramagnetic resonance].
Viscosity, temperature and ionic strength dependences of ESR microwave saturation parameters of spin labelled human oxyhemoglobin (Hb) and bovine serum albumin (BSA) have been studied. The piperidine and pyrrolidine nitroxyl derivatives of maleimide were used as covalent SH reagents for Hb and BSA and the same two derivatives of gamma-benzocarboline and spin labelled stearic acid were used as noncovalent spin probes for BSA. The effects of label binding tightness on ESR spectral parameters were considered. The rotational correlation times were determined using viscosity dependences of the separation of the outer hyperfine extrema and Stokes extrapolations at high viscosities. The ESR microwave saturation parameters of the spin labels were shown to depend just weakly on temperature (at constant eta/t) over the range 0-25 degrees and on g, A values but to be sensitive to protein rotational correlation times up to 10(-4) sec and also to the rotational anisotropy and to the relative motion of the spin label.


Citizen Science in Human Medicine and the Use of Software-Systems: A Rapid Scoping Review.
Citizen science allows involving interested citizen in the entire research process in science. In the past, various citizen science projects have been performed in different research fields, especially in human medicine. We conducted a rapid scoping review to determine which citizen projects in human medicine already used software-based systems to engage citizens in the research process. Furthermore, we analysed which of the software-systems are publicly available, especially in the field of rare diseases, how citizens can participate using those tools and whether the usability was rated by the participants. To get insights for our project "SelEe (Seltene Erkrankungen bürgerwissenschaftlich erforschen)", which is a citizen science project in rare diseases funded by the Federal Ministry of Education and Research (BMBF), we aimed to identify projects in this research area. We searched PubMed for articles between 2011 and 2021 and performed a title- and abstract screening, as well as a full-text screening. Finally, 12 studies were identified in different research areas like public health, genetic research and infectious diseases. We could not identify any study directly associated with rare diseases. None of the studies investigated usability of those systems. Furthermore, five publicly available citizen science software-systems were identified. Three of them are general systems that allow creating, operating, managing citizen science projects and including citizens in the research process. In further investigations, we will check and compare these systems, if they are appropriate for use in our SelEe-project.


Sclerosing cholangitis.
Primary sclerosing cholangitis is a chronic cholestatic liver disease characterized by strictures of the biliary tree complicated by cirrhosis and cholangiocarcinoma. It is immune mediated although the precise etiology remains unknown. Research into etiopathogenesis, epidemiology, the relationship with inflammatory bowel disease, diagnosis of cholangiocarcinoma, medical therapy, and the outcome of liver transplantation are discussed. It is likely that a number of patients previously diagnosed with primary sclerosing cholangitis have autoimmune pancreatitis in association with primary sclerosing cholangitis, a syndrome with distinct clinicopathological features including steroid responsiveness. Primary sclerosing cholangitis-inflammatory bowel disease probably represents a distinct inflammatory bowel disease phenotype, which has implications for colonoscopic surveillance of these patients. CA19-9 plays no surveillance role for the early detection of cholangiocarcinoma. The best-studied drug in primary sclerosing cholangitis is ursodeoxycholic acid, which, despite a range of potentially valuable actions on the cholestatic liver, has not yet been proved to make a substantial impression on the course of the disease. Orthotopic liver transplantation remains the only established long-term treatment for primary sclerosing cholangitis.


Dose response of continuous subcutaneous infusion of recombinant glucagon-like peptide-1 in combination with metformin and sulphonylurea over 12 weeks in patients with type 2 diabetes mellitus.
Any differences observed between natural glucagon-like peptide-1 (GLP-1) and studies obtained with analogues might call for renewed considerations concerning the use and design of such analogues. Thus, we aimed to evaluate the dose-response relationship of recombinant glucagon-like peptide-1 (7-36) amide (rGLP-1) administered by continuous subcutaneous infusion (CSCI) in subjects with type 2 diabetes. We compared the efficacy and safety of three doses of recombinant GLP-1, ranging from 1.25 to 5.0 pmol/kg/min (pkm) and placebo, given by continuous subcutaneous infusion over 3 months in combination with metformin and sulphonylurea (SU), to lower haemoglobin A1c (HbA1c), fasting plasma glucose and weight in 95 type 2 diabetes patients with inadequate glycaemic control. The mean decreases in HbA1c at endpoint (week 12) were significantly greater for all three rGLP-1 dose groups when each was compared with the placebo group, with the greatest decrease occurring in the 5.0 pkm dose group (-1.3%, s.d. ± 0.18, p < 0.001). The mean decreases in fasting plasma glucose from baseline to endpoint were significantly greater for all three rGLP-1 dose groups than for the placebo group, with the greatest decrease occurring in the 5.0 pkm dose group (-26.0 mg/dl, s.d. ± 8.5, p = 0.02). Body weight was significantly reduced by 1.8 kg (s.d. ± 1.3) in the 1.25 pkm dose group only (p = 0.04). Administration of rGLP-1 by CSCI over a 12-week period in combination with metformin and an SU had a dose dependent effect in lowering HbA1c and fasting plasma glucose. However, administration of rGLP-1 by CSCI may be less effective with respect to lowering of body weight compared with the daily and once weekly analogues.


Blood pressure and risk of all-cause mortality in advanced chronic kidney disease and hemodialysis: the chronic renal insufficiency cohort study.
Studies of hemodialysis patients have shown a U-shaped association between systolic blood pressure (SBP) and mortality. These studies have largely relied on dialysis-unit SBP measures and have not evaluated whether this U-shape also exists in advanced chronic kidney disease, before starting hemodialysis. We determined the association between SBP and mortality at advanced chronic kidney disease and again after initiation of hemodialysis. This was a prospective study of Chronic Renal Insufficiency Cohort participants with advanced chronic kidney disease followed through initiation of hemodialysis. We studied the association between SBP and mortality when participants (1) had an estimated glomerular filtration rate <30 mL/min/1.73 m2 (n=1705), (2) initiated hemodialysis and had dialysis-unit SBP measures (n=403), and (3) initiated hemodialysis and had out-of-dialysis-unit SBP measured at a Chronic Renal Insufficiency Cohort study visit (n=326). Cox models were adjusted for demographics, cardiovascular risk factors, and dialysis parameters. A quadratic term for SBP was included to test for a U-shaped association. At advanced chronic kidney disease, there was no association between SBP and mortality (hazard ratio, 1.02 [95% confidence interval, 0.98-1.07] per every 10 mm Hg increase). Among participants who started hemodialysis, a U-shaped association between dialysis-unit SBP and mortality was observed. In contrast, there was a linear association between out-of-dialysis-unit SBP and mortality (hazard ratio, 1.26 [95% confidence interval, 1.14-1.40] per every 10 mm Hg increase). In conclusion, more efforts should be made to obtain out-of-dialysis-unit SBP, which may merit more consideration as a target for clinical management and in interventional trials.


Bioequivalence trials of rifampicin containing formulations: extrinsic and intrinsic factors in the absorption of rifampicin.
Rifampicin shows variable bioavailability from solid oral dosage forms and the reasons for this variable absorption reported in literature varies from extrinsic formulations factors to intrinsic variability in rifampicin absorption. Hence, we have undertaken a systematic and comprehensive evaluation of all the factors to study contribution of all the factors on rifampicin absorption. As a first step, data from eight bioequivalence studies conducted at National Institute of Pharmaceutical Education and Research (NIPER) bioavailability center was compared across the trials to understand the effect of extrinsic/intrinsic factors on the bioavailability of rifampicin, isoniazid and pyrazinamide. Out of eight fixed dose combination (FDC) formulations, six formulations were bioequivalent for rifampicin to separate formulations whereas one formulation was below and one was above the limits of bioequivalence. It was observed that more variability in rifampicin blood levels is associated with FDC formulations when compared to rifampicin-only formulations and was attributed to complexity involved in the manufacturing of FDCs. Further, one of the rifampicin-only capsule showed unexpectedly lower plasma levels indicating role of physical characteristics of rifampicin bulk material. It was also seen that rifampicin shows dose-dependent pharmacokinetics even at the modest increase in dose due to saturation of efflux system at absorption site and metabolizing enzymes for elimination. Other components of FDC formulations such as isoniazid and pyrazinamide due to high solubility and permeability have shown very less variability and were bioequivalent to separate formulations even from formulations those were failed for rifampicin. The comparison of data across the trials suggested that rifampicin bioavailability problem is more attributable to the extrinsic factors such as formulation or rifampicin bulk material rather than intrinsic variability of rifampicin absorption.


Effects of visual map complexity on the attentional processing of landmarks.
In the era of smartphones, route-planning and navigation is supported by freely and globally available web mapping services, such as OpenStreetMap or Google Maps. These services provide digital maps, as well as route planning functions that visually highlight the suggested route in the map. Additionally, such digital maps contain landmark pictograms, i.e. representations of salient objects in the environment. These landmark representations are, amongst other reference points, relevant for orientation, route memory, and the formation of a cognitive map of the environment. The amount of visible landmarks in maps used for navigation and route planning depends on the width of the displayed margin areas around the route. The amount of further reference points is based on the visual complexity of the map. This raises the question how factors like the distance of landmark representations to the route and visual map complexity determine the relevance of specific landmarks for memorizing a route. In order to answer this question, two experiments that investigated the relation between eye fixation patterns on landmark representations, landmark positions, route memory and visual map complexity were carried out. The results indicate that the attentional processing of landmark representations gradually decreases with an increasing distance to the route, decision points and potential decision points. Furthermore, this relation was found to be affected by the visual complexity of the map. In maps with low visual complexity, landmark representations further away from the route are fixated. However, route memory was not found to be affected by visual complexity of the map. We argue that map users might require a certain amount of reference points to form spatial relations as a foundation for a mental representation of space. As maps with low visual complexity offer less reference points, people need to scan a wider area. Therefore, visual complexity of the area displayed in a map should be considered in navigation-oriented map design by increasing displayed margins around the route in maps with a low visual complexity. In order to verify our assumption that the amount of reference points not only affects visual attention processes, but also the formation of a mental representation of space, additional research is required.


Physiological Responses and User Feedback on a Gameful Breathing Training App: Within-Subject Experiment.
Slow-paced breathing training (6 breaths per minute [BPM]) improves physiological and psychological well-being by inducing relaxation characterized by increased heart rate variability (HRV). However, classic breathing training has a limited target group, and retention rates are very low. Although a gameful approach may help overcome these challenges, it is crucial to enable breathing training in a scalable context (eg, smartphone only) and ensure that they remain effective. However, despite the health benefits, no validated mobile gameful breathing training featuring a biofeedback component based on breathing seems to exist. This study aims to describe the design choices and their implementation in a concrete mobile gameful breathing training app. Furthermore, it aims to deliver an initial validation of the efficacy of the resulting app. Previous work was used to derive informed design choices, which, in turn, were applied to build the gameful breathing training app Breeze. In a pretest (n=3), design weaknesses in Breeze were identified, and Breeze was adjusted accordingly. The app was then evaluated in a pilot study (n=16). To ascertain that the effectiveness was maintained, recordings of breathing rates and HRV-derived measures (eg, root mean square of the successive differences [RMSSDs]) were collected. We compared 3 stages: baseline, standard breathing training deployed on a smartphone, and Breeze. Overall, 5 design choices were made: use of cool colors, natural settings, tightly incorporated game elements, game mechanics reflecting physiological measures, and a light narrative and progression model. Breeze was effective, as it resulted in a slow-paced breathing rate of 6 BPM, which, in turn, resulted in significantly increased HRV measures compared with baseline (P<.001 for RMSSD). In general, the app was perceived positively by the participants. However, some criticized the somewhat weaker clarity of the breathing instructions when compared with a standard breathing training app. The implemented breathing training app Breeze maintained its efficacy despite the use of game elements. Moreover, the app was positively perceived by participants although there was room for improvement.


Prenatal diagnosis of fetal ventriculomegaly: Agreement between fetal brain ultrasonography and MR imaging.
Accurate measurement of the lateral ventricles is of paramount importance in prenatal diagnosis. Possible conflicting classifications caused by their measurement in different sectional planes by sonography and MR imaging are frequently raised. The objective of our study was to evaluate the agreement between ultrasonography and MR imaging in the measurement of the lateral ventricle diameter in the customary sectional planes for each technique. Measurement of both lateral ventricles was performed prospectively in 162 fetuses from 21 to 40 weeks of gestational age referred for evaluation due to increased risk for cerebral pathology. The mean gestational age for evaluation was 32 weeks. The measurements were performed in the customary plane for each technique: axial plane for sonography and coronal plane for MR imaging. The 2 techniques yielded results in substantial agreement by using intraclass correlation and κ coefficient score tests. When we assessed the clinical cutoff of 10 mm, the κ score was 0.94 for the narrower ventricle and 0.84 for the wider ventricle, expressing almost perfect agreement. The Bland-Altman plot did not show any trend regarding the actual width of the ventricle, gestational week, or interval between tests. Findings were independent for fetal position, sex, and indication for examination. Our study indicates excellent agreement between fetal brain ultrasonography and MR imaging as to the diagnosis of fetal ventriculomegaly in the customarily used sectional planes of each technique.


Does video-assisted thoracoscopic pleurectomy result in better outcomes than open pleurectomy for primary spontaneous pneumothorax?
The question addressed by a best evidence topic approach using a structured protocol was whether pleurectomy using video-assisted thoracoscopic surgery (VATS) resulted in better outcomes than open pleurectomy for primary spontaneous pneumothorax. Altogether 45 relevant papers were identified of which nine papers represented the best evidence to answer the question. The author, journal, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses were tabulated. We conclude that VATS pleurectomy has been shown to be comparable to open pleurectomy in the treatment of spontaneous pneumothorax, with a meta-analysis and several RCTs showing reductions in length of hospital stay and analgesic requirements. Postoperative pulmonary dysfunction has also been shown to be reduced after VATS pleurectomy in two RCTs, although a third study found no significant difference. A concern may be a four-fold increase in the recurrence of pneumothorax following VATS pleurectomy as compared to open pleurectomy reported in a recent meta-analysis of four randomised and 25 non-randomised studies performed in 2007 and published in the Lancet, although a second meta-analysis of only the randomised trials did not show this difference.


Protein and vitamin B6 intake are associated with liver steatosis assessed by transient elastography, especially in obese individuals.
Although the detrimental effects of several dietary components on the promotion of nonalcoholic fatty liver disease are well known, no studies have assessed the role of dietary vitamin B6. Moreover, studies on the associations between dietary components or body composition indices and liver steatosis assessed by transient elastography are rare. Our aim was to identify the nutritional factors and anthropometric parameters associated with liver steatosis. In this cross-sectional study, we enrolled 168 individuals (35% obese) who underwent a liver steatosis assessment by Controlled Attenuation Parameter measurement and nutritional assessment. Tertiles of vitamin B6 intake were positively associated with hepatic steatosis (B=1.89, P=0.026, confidence interval [CI] 0.03-0.80) as well as with triglycerides, glucose, alanine aminotransferase (ALT), and body mass index . In obese individuals, after multivariable analysis, the Controlled Attenuation Parameter score was still associated with triglycerides, ALT, and total protein intake (B=0.56, P=0.01, CI 0.10-1.02). Participants in tertile I (low intake) had a lower Controlled Attenuation Parameter than those in tertile III (P=0.01). We found a positive association between hepatic steatosis or Controlled Attenuation Parameter score and vitamin B6/total protein intake, probably related to the high intake of meat. Vitamin B6 might have a pathogenic role related to the increase of hepatic steatosis.


Complications from tourniquet use.
Complications of tourniquet use are rare, but when they do occur, they may be devastating. This article outlines potential complications of tourniquet use and recommends measures that can be taken by clinicians using this valuable and potentially dangerous tool.


Clinical and diagnostic imaging findings in an Italian wolf (Canis lupus italicus) with discospondylitis.
An adult male Italian wolf (Canis lupus italicus) was presented with an abnormal gait. Neurologic examination showed thoracic kyphosis, paraparesis, decreased proprioception in the pelvic limbs, and normal spinal reflexes. Neurologic symptoms suggested a thoracolumbar spinal cord lesion. Pathologic findings included leukocytosis. Spinal radiographs revealed ventral spondylosis of T4/T5/T6, a poorly defined intervertebral disc space, and mild lysis of the vertebral margins. Multiple metallic foreign bodies were seen in the thoracic wall. Magnetic resonance imaging of the spine detected increased signal intensity on fluid sensitive sequences of the vertebral bodies, the intervertebral disc, and surrounding soft tissues. These findings were interpreted as active discospondylitis at T4/T5. Medical therapy included antibiotic and analgesic treatment as well as movement restriction. Follow-up at 4 wk showed significant clinical and radiologic improvement. Discospondylitis should be included in the differential diagnosis in wolves with paresis.


Temperature shapes language sonority: Revalidation from a large dataset.
Multiple factors of the natural environment have been found to impact and mold the phonetic patterns of human speech, among which the potential correlation between sonority and temperature has garnered significant attention. We leverage a large database containing basic vocabularies of 5,293 languages and calculate the average sonority for each language by adopting a universal sonority scale. Our findings confirm a positive correlation between sonority and temperature across macroareas and language families, whereas this relationship cannot be discerned within language families. We suggest that the adaptation of the distribution of speech sounds within languages is a slow process which is moreover insensitive to minor differences in temperature experienced by speakers as they carry their languages to new regions. Nevertheless, at the global level a solid relationship emerges. Furthermore, we delve deeper into the nature of the relationship and contend that it is mainly due to cold temperatures having a weakening effect on sonority. This research provides compelling additional evidence that climatic factors contribute to shaping language and its evolution.


Evidence regarding cosmetic and medically unnecessary surgery on infants.
The Journal of Pediatric Urology has recently published several articles from the Annecy (France) Working Party on DSD. We question several of the presented findings and recommendations. In two key articles summarizing their review, the authors concluded that identified studies are not representative and suffer from methodological weaknesses, such that they "lack the necessary detail to base further recommendations". In a third article, the Working Party reported that the science supporting early surgery is "scanty", and that "no studies" support the belief that gender variant children require early genital surgery. Nevertheless, the Working Party warned that without long-term research, "if no effort is made, we will be left, in the next generation, to continue making the same judgment, based on 'experience' and 'expert opinion'". None of the studies cited in the articles support such assertions as we read them. We maintain that reviewed evidence suggests a moratorium on early surgical intervention is imperative for children with differences in sex development, and that the best ethical and scientific considerations require that gender surgery should be delayed until the child can consent. We further present evidence that UN and case law presently under way in the USA support such a moratorium.


Barriers and Facilitators of Quality Family-Centered Communication in Pakistan.
Communication is a fundamental aspect of patient- and family-centered care. Unfortunately, there is a dearth of evidence regarding pediatric cancer communication in low- and middle-income countries, where over 90% of all children with childhood cancer live. The purpose of this study was to explore barriers and facilitators of quality communication within two pediatric cancer centers in Pakistan. Semistructured interviews were conducted with 20 multidisciplinary pediatric cancer clinicians and 18 caregivers of children with cancer at Children's Hospital of Lahore and Indus Hospital in Karachi, Pakistan. Interviews were conducted in English or Urdu, audio-recorded, transcribed, and translated to English. Two researchers coded each transcript using an inductively derived codebook. Thematic content analysis focused on barriers and facilitators of high-quality communication. Pakistani clinicians and caregivers identified factors that affected the quality of patient-centered cancer communication. These included structural factors including setting, available interpreters, documentation, patient volume, teamwork, and financial support. Clinician-level communication barriers and facilitators included communication training, clinician distress/boundaries, and the ability to have recurrent conversations. Patient or family characteristics affecting communication included education, income status, primary language, and geography; the child's specific disease type; and relational elements such as social support, empowerment, and split decision makers. Participants identified existing or potential interventions related to each factor. Multilevel factors serve as either barriers or facilitators for pediatric cancer communication in Pakistan. Identification of these elements of communication is an essential step toward interventions aimed at improving patient- and family-centered care in resource limited settings.


The Effect of Thyroid Eye Disease on Corneal Biomechanical Properties.
The aim of this study was to identify corneal biomechanical parameters measured by ORA in patients with TED compared to the healthy group. The NOSPECS classification of patients is used to assess the relation between biomechanical changes and disease severity. We included 22 TED patients, diagnosed with TED for more than five years, and 43 healthy participants. The NOSPECS classification was assessed as mild (grade 1-3) and severe (grade 4-6) disease. For each group, corneal hysteresis (CH), corneal resistance factor (CRF), central corneal thickness (CCT), Goldmann-correlated intraocular pressure (IOPg) and corneal compensated intraocular pressure (IOPcc) parameters were measured by ORA. The mean age was 38.8±11.6 years for the TED patients and 42.9±15.58 years for the control group. For TED patients and healthy volunteers, the mean levels of CRF, CH, and CCT were measured as follows: 10.43±2.04 vs 10.28±1.91mmHg, p=0.67; 10.18±1.81 vs 10.21±1.68 mmHg, p=0.90; 550.31±35.73 vs 545.23±37.91 µm, p=0.47, respectively. These values were not significant between groups, but they were significantly higher in females compared to males in TED patients [CRF;10.68 (IQR: 9.49-12.14) vs 8.96 (IQR: 8.04-9.92) mmHg, p=0.002, CH; 10.43 (IQR: 9.48-11.25) vs 8.58 (IQR: 7.90-9.95) mmHg, p=0.003 and CCT; 554.25 (IQR: 536.05-579.52) vs 527.40 (IQR: 492.25-545.90) μm, p=0.014]. CRF values were negatively correlated with NOSPECS score (r=-0.317, p=0.036) and significantly higher CRF was observed in mild patients compared to severe disease (11.43 (IQR: 10.14-12.87) vs 9.46 (IQR: 8.75-10.28) mmHg, p=0.008). We found a significant gender effect on corneal biomechanical parameters of TED patients. CRF, CH and CCT values were significantly higher in females compared to males with TED. The clinical severity score of TED showed negative correlation with CRF. CRF value might be a useful parameter in follow-up of TED patients in clinical practice.


The impact of cigarette smoking on sleep quality of patients with masticatory myofascial pain.
To evaluate the impact of cigarette smoking on the sleep quality of patients with masticatory myofascial pain and to determine whether the association between smoking and impaired sleep is influenced by other factors such as demographic, psychological, and behavioral variables. Data from a retrospective case series of 529 patients diagnosed with masticatory myofascial pain according to group I of the Research Diagnostic Criteria for Temporomandibular Disorders were obtained. Patients completed a standardized pain questionnaire and psychometric tests. Differences between smokers and nonsmokers were evaluated using independent samples t tests and chi-square tests. Hierarchical linear multiple regression models were used to examine predictors of sleep disturbances. Of the entire sample, 170 (32%) reported that they were smokers. Smokers reported higher pain severity and more sleep disturbances and psychological distress than nonsmokers. Cigarette smoking significantly predicted sleep disturbance (β = 0.229, P < .001), but this relationship was attenuated after controlling for pain severity and psychological distress (β = 0.122, P < .001). Cigarette smoking is associated with numerous adverse health outcomes, including pain severity, alterations in mood, and disrupted sleep, and seems to be a significant predictor of sleep quality in patients with masticatory myofascial pain.


Bile salts induce long polar fimbriae expression favouring Crohn's disease-associated adherent-invasive Escherichia coli interaction with Peyer's patches.
Ileal lesions of patients with Crohn's disease are colonized by adherent-invasive Escherichia coli (AIEC). The earliest lesions of recurrent Crohn's disease are erosions of Peyer's patches (PP). We recently reported the presence of a functional lpf operon in AIEC, encoding long polar fimbriae (LPF), that allows AIEC bacteria to interact with PP and to translocate across M cells. The aim of this study was to analyse the effect of gastrointestinal conditions on LPF expression in AIEC strains. The LF82 bacterial growth in an acid pH medium or at high osmolarity medium had no effect on lpf transcription level, in contrast to bacterial growth in the presence of bile salts, which promoted activation of lpf transcription. When cultured in the presence of bile salt, LF82 wild-type bacteria, but not the isogenic mutant deleted for lpfA, exhibited a higher level of interaction with PP and a higher level of translocation through M cell monolayers. The FhlA transcriptional factor was found to be a key bacterial regulator at the origin of LPF expression in the presence of bile salts.


Methionine and leucine enkephalin in rat neurohypophysis: different responses to osmotic stimuli and T2 toxin.
Specific radioimmunoassays were used to measure the effects of hypertonic saline (salt loading), water deprivation, and trichothecene mycotoxin (T2 toxin) on the content of methionine enkephalin (ME), leucine enkephalin (LE), alpha-neoendorphin, dynorphin A, dynorphin B, vasopressin, and oxytocin in the rat posterior pituitary. Concentrations of vasopressin and oxytocin decreased in response to both osmotic stimuli and treatment with T2 toxin, but the decrease was greater with osmotic stimulations. Similarly, concentrations of LE and dynorphin-related peptides declined after salt loading and water deprivation; LE concentrations also decreased after treatment with T2 toxin. The concentration of ME decreased after water deprivation, did not change after salt loading, and increased after T2 toxin treatment. The differentiating effects of these stimuli on the content of immunoreactive LE and ME are consistent with the hypothesis that LE and ME may be localized in separate populations of nerve endings with different roles in the posterior pituitary.


Zn(ii)-Curcumin supplementation alleviates gut dysbiosis and zinc dyshomeostasis during doxorubicin-induced cardiotoxicity in rats.
Doxorubicin is a powerful anticancer agent used to treat a variety of human neoplasms. However, the clinical use of doxorubicin is hampered by cardiotoxicity and effective cardioprotective adjuvants do not exist. Dietary zinc, an essential nutrient, is required to maintain steady-state tissue zinc levels and intestinal homeostasis and may yield therapeutic benefits in diseases associated with zinc dysregulation or gut dysbiosis. Here, we investigated the effects of dietary Zn(ii)-curcumin (ZnCM) solid dispersions on gut dysbiosis and zinc dyshomeostasis during doxorubicin-induced cardiotoxicity in rats. Rats were injected with multiple low doses of doxorubicin and orally administered ZnCM daily over four weeks. Daily administration of ZnCM not only alleviated Dox-induced gut dysbiosis-as indicated by the increased Firmicutes-to-Bacteroidetes ratio and the maintenance of the relative abundances of major beneficial bacteria including Clostridium_XIVa, Clostridium_IV, Roseburia, Butyricicoccus and Akkermansia-but also maintained intestinal barrier integrity and decreased the lipopolysaccharide (LPS) contents of feces and plasma. ZnCM also significantly attenuated doxorubicin-induced zinc dyshomeostasis, which was mirrored by preservation of zinc levels and expression of zinc-related transporters. Furthermore, ZnCM significantly improved heart function and reduced cardiomyocyte apoptosis and myocardial injury in doxorubicin-treated rats. Notably, the regulation of zinc homeostasis and cardioprotective and microbiota-modulating effects of ZnCM were transmissible through horizontal feces transfer from ZnCM-treated rats to normal rats. Thus, ZnCM supplementation has potential as an effective therapeutic strategy to alleviate gut dysbiosis and zinc dyshomeostasis during doxorubicin-induced cardiotoxicity.


Serum cholesterol and triglyceride levels in 3,446 children from a biracial community: the Bogalusa Heart Study.
Serum lipid profiles of 3,446 (91% of population) children, ages 5-14 years, were determined in a biracial community (Bogalusa, Louisiana) as part of a program investigating the early natural history of atherosclerosis. Black children had significantly higher mean levels of serum cholesterol than white children (170 mg/dl vs 162 mg/dl, P less than 0.0001). On the other hand, significantly lower levels of triglycerides were found in blacks than in whites (61 mg/dl vs 73 mg/dl, P less than 0.0001). Girls had higher levels of triglycerides than boys in both races (blacks, 64 mg/dl vs 59 mg/dl, P less than 0.001; whites, 77 mg/dl vs 69 mg/dl, P less than 0.001). The racial differences in serum cholesterol and triglyceride levels were even more apparent at the 95th percentile. The serum cholesterol level remained relatively constant in all children until ages 11 and 12 years, after which a slight reduction occurred. This reduction was more pronounced in boys than in girls. In contrast, a significant increase in the level of triglycerides with age was observed in all children except black girls, the increasing slope being most pronounced in white girls.


High response rate in refractory and poor-risk multiple myeloma after allotransplantation using a nonmyeloablative conditioning regimen and donor lymphocyte infusions.
Standard allogeneic stem cell transplant (allo-SCT) regimens have been associated with a high transplant-related mortality (TRM) in multiple myeloma (MM). Nonmyeloablative therapy can establish stable engraftment after allo-SCT and maintain the antitumor effect with less toxicity, which is important in heavily pretreated and elderly patients. We report on 16 poor-risk MM patients receiving allo-SCT from an HLA-matched (n = 14) or mismatched (n = 2) sibling following conditioning with melphalan 100 mg/m(2) (MEL-100). Ten patients had refractory relapse, 4 responsive relapse, and 2 patients were in near complete remission (nCR) with poor-prognosis disease. Patients had received 1 (n = 9) or 2 (n = 7) prior autotransplants. Donor lymphocyte infusions (DLIs) were given to 14 patients with no clinical evidence of graft versus host disease (GVHD) either to attain full donor chimerism (n = 4) or to eradicate residual disease (n = 10). Fifteen patients showed myeloid engraftment, and 12 patients were full donor chimeras at day +21. No TRM was observed during the first 100 days. Acute GVHD developed in 10 patients; 1 had fatal grade IV GVHD. Seven progressed to chronic GVHD, limited in 3 and extensive in 4 patients. At a median follow-up of 1 year, 5 patients achieved and sustained CR, 3 nCR, and 4 partial remission. Of 4 patients progressing after transplantation, 3 achieved a remission following further chemotherapy and DLI. Remarkable graft versus myeloma responses were seen in chemotherapy-refractory patients. Two patients died of progressive disease, and 3 died of GVHD complications without active disease. GVHD remains a major problem with this procedure.


Comparison of the appearance of radiolabelled vitamin D3 and 25-hydroxy-vitamin D3 in the chylomicron fraction of plasma after oral administration in man.
1. The uptake of either orally administered [3H]vitamin D3 or 25-[3H]hydroxy-vitamin D3 into the chylomicron fraction of plasma was studied in 12 healthy male subjects. 2. The amount of [3H]vitamin D3 in the chylomicron fraction expressed as a percentage of the total plasma radioactivity was higher than that of 25-[3H]hydroxy-vitamin D3. 3. It is suggested that the mechanism of intestinal absorption of vitamin D and 25-hydroxy-vitamin D may differ in man, the absorption of 25-hydroxy-vitamin D possibly being less dependent on bile acids.


The limits of physiological anemia in the African neonate.
In order to determine the limits of physiological anemia in the African neonate, the hematological values of 402 healthy neonates were determined. It was observed that although the African neonate had much lower hematological values at birth when compared with their North American and European counterparts, the eventual nadir reached is approximately the same. The hematocrit, hemoglobin and red blood cell count of preterm infants on the first day of life were comparable to those of full-term and postterm infants. However, the preterm infants had a more precipitous drop in these values and, consequently, reached a nadir much lower than full-term and postterm infants. The nadir was reached between 6 and 8 weeks in each group. In all three groups, recovery was spontaneous and preceded by a rise in reticulocyte count. It is also significant that by 16 weeks of life, no significant difference exists between the values for preterm, full-term and postterm infants.


Timing of physical activity within the 24-hour day and its influence on health: a systematic review.
Recent studies report that the health benefits of physical activity differ depending on whether the activity is performed in the morning, afternoon or evening. The purpose of this systematic review was to examine whether the timing of physical activity within the 24-hour day is associated with health. Five databases were searched for English or French language peer-reviewed studies that examined whether the timing of physical activity within the day is associated with health. No limits were placed on publication year, study population, study design or health outcomes. Studies that examined acute effects of physical activity or timing of physical activity around food intake were excluded. This systematic review examined 35 studies, with 17 259 participants, and the following health outcomes: measures of sleep health, adiposity, fat-free mass and muscle size, cardiometabolic biomarkers, physical function and mobility, mental health, and risk of cardiovascular disease, cancer, and mortality. Heterogeneity across studies precluded meta-analyses, and we present our findings using narrative syntheses. Of the 35 studies, 11 reported that morning physical activity provides greater health benefits than afternoon/ evening physical activity, while 12 found that morning physical activity provides fewer health benefits than afternoon/evening physical. In the remaining 12 studies, there was no clear difference in health benefits based on the timing of physical activity. The quality of evidence for the different health outcomes across study designs was very low. There is no consistent evidence that physical activity at one time of day provides more favourable health benefits than physical activity at a different time of day. (PROSPERO registration no.: CRD42021231088).


Molecularly imprinted covalent organic polymers for the selective extraction of benzoxazole fluorescent whitening agents from food samples.
Molecularly imprinted covalent organic polymers were constructed by an imine-linking reaction between 1,3,5-triformylphloroglucinol and 2,6-diaminopyridine and used for the selective solid-phase extraction of benzoxazole fluorescent whitening agents from food samples. Binding experiments showed that imprinting sites on molecularly imprinted polymers had higher selectivity for targets compared with those of the corresponding non-imprinted polymers. Parameters affecting the solid-phase extraction procedure were examined. Under optimal conditions, actual samples were treated and the eluent was analyzed with high-performance liquid chromatography with diode-array detection. The results showed that the established method has a wide linearity, satisfactory detection limits and quantification limits, and acceptable recoveries. Thus, this developed method possesses the practical potential for the selective determination of benzoxazole fluorescent whitening agents in complex food samples.


Intracardiac thrombosis and embolism in patients with cardiac amyloidosis.
Patients with primary amyloidosis (AL type) have a poor prognosis, in part due to frequent cardiac involvement. Although intracardiac thrombus has been reported in anecdotal cases, neither its frequency nor its role in causing mortality is known. Furthermore, the clinical and echocardiographic variables that may be associated with thromboembolism in cardiac amyloidosis have not been defined. A total of 116 autopsy or explanted cases of cardiac amyloidosis (55 AL and 61 other type) were identified in the Mayo Clinic. Forty-six fatal nonamyloid trauma cases served as controls. Each heart was examined for intracardiac thrombus. The cause of death was determined from autopsy and clinical notes. Intracardiac thrombosis was identified in 38 hearts (33%). Twenty-three had 1 thrombus, whereas 15 had 2 to 5 thrombi. Although subjects in the AL group were younger and had less atrial fibrillation than those with other types of amyloidosis, the AL group had significantly more intracardiac thrombus (51% versus 16%, P<0.001) and more fatal embolic events (26% versus 8%, P<0.03). Control hearts had no intracardiac thrombus. The presence of both atrial fibrillation and AL was associated with an extremely high risk for thromboembolism (odds ratio 55.0 [95% confidence interval 8.1 to 1131.4]). By multivariate analysis, AL type (odds ratio 8.4 [95% confidence interval 1.8 to 51.2]) and left ventricular diastolic dysfunction (odds ratio 12.2 [95% confidence interval 2.7 to 72.7]) were independently associated with thromboembolism. A high frequency of intracardiac thrombosis was present in cardiac amyloidosis. Furthermore, thromboembolism caused significant fatality. Several risk factors for thromboembolism were identified. Early screening, especially in high-risk patients, and early anticoagulation might reduce morbidity and mortality.


A new genetic locus for antipsychotic-induced weight gain: A genome-wide study of first-episode psychosis patients using amisulpride (from the OPTiMiSE cohort).
Antipsychotic-induced weight gain is a common and debilitating side effect of antipsychotics. Although genome-wide association studies of antipsychotic-induced weight gain have been performed, few genome-wide loci have been discovered. Moreover, these genome-wide association studies have included a wide variety of antipsychotic compounds. We aim to gain more insight in the genomic loci affecting antipsychotic-induced weight gain. Given the variable pharmacological properties of antipsychotics, we hypothesized that targeting a single antipsychotic compound would provide new clues about genomic loci affecting antipsychotic-induced weight gain. All subjects included for this genome-wide association study (n=339) were first-episode schizophrenia spectrum disorder patients treated with amisulpride and were minimally medicated (defined as antipsychotic use <2 weeks in the previous year and/or <6 weeks lifetime). Weight gain was defined as the increase in body mass index from before until approximately 1 month after amisulpride treatment. Our genome-wide association analyses for antipsychotic-induced weight gain yielded one genome-wide significant hit (rs78310016; β=1.05; p=3.66 × 10-08; n=206) in a locus not previously associated with antipsychotic-induced weight gain or body mass index. Minor allele carriers had an odds ratio of 3.98 (p=1.0 × 10-03) for clinically meaningful antipsychotic-induced weight gain (⩾7% of baseline weight). In silico analysis elucidated a chromatin interaction with 3-Hydroxy-3-Methylglutaryl-CoA Synthase 1. In an attempt to replicate single-nucleotide polymorphisms previously associated with antipsychotic-induced weight gain, we found none were associated with amisulpride-induced weight gain. Our findings suggest the involvement of rs78310016 and possibly 3-Hydroxy-3-Methylglutaryl-CoA Synthase 1 in antipsychotic-induced weight gain. In line with the unique binding profile of this atypical antipsychotic, our findings furthermore hint that biological mechanisms underlying amisulpride-induced weight gain differ from antipsychotic-induced weight gain by other atypical antipsychotics.


BRI2 interacts with amyloid precursor protein (APP) and regulates amyloid beta (Abeta) production.
Transmembrane proteins BRI2 and amyloid precursor protein (APP) co-localize with amyloid beta (Abeta) lesions in sporadic Alzheimer disease and mutations in both precursor proteins are linked to early-onset familial cases of cerebral amyloidosis associated with dementia and/or cerebral hemorrhage. A specific interaction between BRI2 and APP was unveiled by immunoprecipitation experiments using transfected and non-transfected cells. The use of deletion mutants further revealed that stretches 648-719 of APP751 and 46-106 of BRI2, both inclusive of the full transmembrane domains, are sufficient for the interaction. Removal of most of the APP and BRI2 extracellular domains without affecting the interaction implies that both proteins interact when are expressed on the same cell membrane (cis) rather than on adjacent cells (trans). The presence of BRI2 had a modulatory effect on APP processing, specifically increasing the levels of cellular APP as well as beta-secretase-generated COOH-terminal fragments while decreasing the levels of alpha-secretase-generated COOH-terminal fragments as well as the secretion of total APP and Abeta peptides. Determining the precise molecular pathways affected by the specific binding between APP and BRI2 could result in the identification of common therapeutic targets for these sporadic and familial neurodegenerative disorders.


[Staphylococci in the skin microbiocenosis of the breasts in pregnant women].
The microflora of the mammary glands in the area of the nipple, the areola and the adjacent skin was studied by the methods of washings and impression. 120 nonpregnant women and 164 pregnant women were examined. The pregnant women showed a higher level of the contamination of the above-mentioned sites. The highest density of bacterial population was detected in the area of the nipple and the lowest density, on the skin surrounding the areola. Coagulase-negative staphylococci proved to be the most numerous organisms among all bacterial population found on the skin of the mammary glands of pregnant women. Of these staphylococci, S. epidermidis was most frequently isolated, its isolation rate being higher in the pregnant women than in the nonpregnant ones.


Activity-Related Outcomes of Articular Cartilage Surgery: A Systematic Review.
The purpose of this systematic review was to compare activity-based outcomes after microfracture, autologous chondrocyte implantation (ACI), and osteochondral autograft (OAT). Multiple databases were searched with specific inclusion and exclusion criteria for level III and higher studies with activity outcomes after microfracture, OAT, osteochondral allograft, and ACI. Activity-based outcomes included the Knee Injury and Osteoarthritis Outcome Score (KOOS), the Tegner Score, the Cincinnati Knee scores, the International Knee Documentation Committee (IKDC) subjective knee score, the Marx activity score, and/or the rate of return-to-sport. Twenty studies were included (1,375 patients). Although results were heterogeneous, significant advantages were seen for ACI and OAT as compared with microfracture in Tegner scores at 1 year (ACI vs. microfracture, P = 0.0016), IKDC scores at 2 years (ACI vs microfracture, P = 0.046), Lysholm scores at 1 year (OAT vs microfracture, P = 0.032), and Marx scores at 2 years (OAT vs microfracture, P < 0.001). The only score or time point to favor microfracture was Lysholm score at 1 year (ACI vs microfracture, P = 0.037). No other standardized outcome measures or time points were significantly different. Several studies demonstrated significantly earlier return to competition with microfracture. Overall reoperation rates were similar, but of reoperations, a higher proportion of those following ACI were unplanned with the majority of performed for graft delamination or hypertrophy. ACI and OAT may have some benefits over microfracture, although return-to-sport is fastest following microfracture. Heterogeneity in technique, outcome measures, and patient populations hampers systematic comparison within the current literature.


Staphylococcus aureus physiological growth limitations: insights from flux calculations built on proteomics and external metabolite data.
Comparing proteomics and metabolomics allows insights into Staphylococcus aureus physiological growth. We update genome and proteome information and deliver strain-specific metabolic models for three S. aureus strains (COL, N315, and Newman). We find a number of differences in metabolism and enzymes. Growth experiments (glucose or combined with oxygen limitation) were conducted to measure external metabolites. Fluxes of the central metabolism were calculated from these data with low error. In exponential phase, glycolysis is active and amino acids are used for growth. In later phases, dehydroquinate synthetase is suppressed and acetate metabolism starts. There are strain-specific differences for these phases. A time series of 2-D gel protein expression data on COL strain delivered a second data set (glucose limitation) on which fluxes were calculated. The comparison with the metabolite-predicted fluxes shows, in general, good correlation. Outliers point to different regulated enzymes for S. aureus COL under these limitations. In exponential growth, there is lower activity for some enzymes in upper glycolysis and pentose phosphate pathway and stronger activity for some in lower glycolysis. In transition phase, aspartate kinase is expressed to meet amino acid requirements and in later phases there is high expression of glyceraldehyde-3-phosphate dehydrogenase and lysine synthetase. Central metabolite fluxes and protein expression of their enzymes correlate in S. aureus.


Analysis of long-term culture properties and pluripotent character of two sibling human embryonic stem cell lines derived from discarded embryos.
We had earlier reported the derivation and characterization of two new sibling human embryonic stem cell lines BJNhem19 and BJNhem20, from discarded grade III embryos of Indian origin. We report here the characteristics of the two sibling cell lines after long-term continuous culture for over 2 yr during which they have been passaged over 200 times. We show that both cell lines adapt well to culture on various mouse and human feeders as well as in feeder-free conditions. The cells show normal diploid karyotype and continue to express all pluripotency markers. Both cell lines differentiate to derivatives of all three germ layers in vitro. However as reported earlier, BJNhem19 is unable to generate teratomas in nude or SCID mice or differentiate to beating cardiomyocytes when tested over several passages during long-term stable culture. On the other hand, the cardiac differentiation capacity of BJNhem20 is greatly increased, and it can generate beating cardiomyocytes that proliferate when isolated and cultured further. In conclusion, the two cell lines have maintained a stable phenotype for over 2 yr and are indeed immortal. Their derivation from grade III embryos does not seem to have any adverse effect on their long-term phenotype. The cells can be obtained for research purposes from the UK Stem Cell Bank and from the authors.


A systematic review of the relationship between polymorphic sites in the estrogen receptor-beta (ESR2) gene and breast cancer risk.
The estrogen signal is mediated by the estrogen receptor (ER). The specific role of ER-beta, a second ER, in breast carcinogenesis is not known. A number of association studies have been carried out to investigate the relationship between polymorphic sites in the ESR2 gene and breast cancer risk, however, the results are inconsistent. We searched PubMed, Medline, and Web of Science database (updated to 10 January 2010) and identified 13 relevant case-control studies, and approximately 28 single-nucleotide polymorphisms (SNPs) and one micro-satellite marker were reported in the literature. The median number of study subjects was 776 (range 158-13,550). Three genetic variants [(CA)n, rs2987983, and rs4986938] showed significant overall associations with breast cancer, and rs4986938 was reported twice. Because rs4986938 and rs1256049 were the most extensively studied polymorphisms, we subsequently conducted a meta-analysis to evaluate their relationship with breast cancer risk (9 studies of 10,837 cases and 16,021 controls for rs4986938; 8 studies of 11,652 cases and 15,726 controls for rs1256049). For rs4986938, the women harboring variant allele seemed to be associated with a decreased risk either in the dominant model [pooled OR = 0.944, 95% confidence interval (95% CI) 0.897-0.993, fixed-effects] or in the co-dominant model (AG vs. GG) (OR = 0.944, 95% CI 0.895-0.997, fixed-effects). rs1256049 was not associated with breast cancer risk in any model. Five studies had investigated the effect of haplotypes in the ESR2 gene on breast cancer risk, and four of them had positive outcomes. In summary, the present systematic review suggests that SNP rs4986938 as well as haplotypes in the ESR2 gene might be associated with breast cancer. The need for additional studies examining these issues seems of vital importance.


Mycobacterium tuberculosis Immune Response in Patients With Immune-Mediated Inflammatory Disease.
Subjects with immune-mediated inflammatory diseases (IMID), such as rheumatoid arthritis (RA), have an intrinsic higher probability to develop active-tuberculosis (TB) compared to the general population. The risk ranges from 2.0 to 8.9 in RA patients not receiving therapies. According to the WHO, the RA prevalence varies between 0.3% and 1% and is more common in women and in developed countries. Therefore, the identification and treatment of TB infection (TBI) in this fragile population is important to propose the TB preventive therapy. We aimed to study the M. tuberculosis (Mtb) specific T-cell response to find immune biomarkers of Mtb burden or Mtb clearance in patients with different TB status and different risk to develop active-TB disease. We enrolled TBI subjects as example of Mtb-containment, the active-TB as example of a replicating Mtb status, and the TBI-IMID as fragile population. To study the Mtb-specific response in a condition of possible Mtb sterilization, we longitudinally enrolled TBI subjects and active-TB patients before and after TB therapy. Peripheral blood mononuclear cells were stimulated overnight with Mtb peptides contained in TB1- and TB2-tubes of the Quantiferon-Plus kit. Then, we characterized by cytometry the Mtb-specific CD4 and CD8 T cells. In TBI-IMID, the TB therapy did not affect the ability of CD4 T cells to produce interferon-γ, tumor necrosis factor-α, and interleukin-2, their functional status, and their phenotype. The TB therapy determined a contraction of the triple functional CD4 T cells of the TBI subjects and active-TB patients. The CD45RA- CD27+ T cells stood out as a main subset of the Mtb-specific response in all groups. Before the TB-preventive therapy, the TBI subjects had higher proportion of Mtb-specific CD45RA-CD27+CD4+ T cells and the active-TB subjects had higher proportion of Mtb-specific CD45RA-CD27-CD4+ T cells compared to other groups. The TBI-IMID patients showed a phenotype similar to TBI, suggesting that the type of IMID and the IMID therapy did not affect the activation status of Mtb-specific CD4 T cells. Future studies on a larger and better-stratified TBI-IMID population will help to understand the change of the Mtb-specific immune response over time and to identify possible immune biomarkers of Mtb-containment or active replication.


Influence of cottonseed meal supplementation on voluntary intake, ruminal and cecal fermentation, digesta kinetics and serum insulin and growth hormone in mature ewes fed prairie hay.
To determine the influence of protein supplementation on intake and fermentation of low-quality hay, six ruminal- and cecal-cannulated Rambouillet ewes (avg wt 43.6 kg) in a crossover design were given ad libitum access to prairie hay with or without 80 g of cottonseed meal (CSM) X head-1 X d-1. Voluntary hay intake was measured the last 7 d of each 18-d period. Ruminal, cecal and blood samples were collected at 0, 1 (except cecal), 3, 6, 9, 12, 15, 18, 21 and 24 h post-supplementation on d 14 of each period to measure fluid dilution rate, fermentation characteristics and serum concentrations of insulin and growth hormone. An intraruminal dose of Yb-labeled hay, followed by fecal sampling on d 15 through 18, was used to measure particulate passage rate. Voluntary intake of prairie hay was increased (P less than .04) from 23.7 to 28.3 g/kg of body weight by CSM supplementation. Particulate passage rate constants did not differ (P greater than .15) between supplemented (3.76%/h) and control (3.72%/h) ewes, and total mean retention time was not altered (P greater than .15) by CSM supplementation. Ruminal retention time of particulates did not differ (P greater than .15) between treatments; however, intestinal transit time was faster (P less than .03; 18.1 vs 22.6 h) in supplemented than in control ewes. Estimated gastrointestinal dry matter fill was greater (P less than .05; 14.3 vs 12.9 g/kg body weight) in supplemented ewes. Ruminal fluid volume did not differ (P greater than .15) between treatments; however, supplemented ewes tended to have faster fluid dilution rates (P less than .14) and fluid outflow rates (P less than .11) than control ewes. Cecal fluid volume, dilution rate and outflow rate did not differ (P greater than .15) between groups. Ruminal and cecal pH and total volatile fatty acids were similar between treatments. Similarly, cottonseed meal supplementation did not affect (P greater than .15) ruminal or cecal ammonia concentrations. Molar proportions of ruminal and cecal individual fatty acids were not affected (P greater than .15) by CSM supplementation. Feeding cottonseed meal increased (P less than .05) serum insulin, decreased (P less than .07) serum growth hormone and increased (P less than .06) serum free fatty acids, but did not influence (P greater than .15) either serum urea N or glucose concentrations. Cottonseed meal supplementation in ewes fed prairie hay caused increased hay intake but had minimal effects on ruminal and cecal fermentation.


Autologous hematopoietic stem cell transplantation is superior to alemtuzumab in patients with highly active relapsing multiple sclerosis and severe disability.
To assess the differences of treatment outcomes regarding disease activity in patients with highly active relapsing multiple sclerosis (RMS), treated with autologous hematopoietic stem cell transplantation (HSCT) or alemtuzumab (ATZ). Open-label prospective single-center observational cohort study, enrolling patients with highly active RMS for treatment with ATZ or HSCT between 2014 and 2021. A total of 50 patients (31/50 (62 %) in HSCT vs 19/50 (38 %) in ATZ group) were included. There were no significant differences in relapse rate, MRI activity or disability worsening between the two study groups during the first two years after treatment onset. However, at 3 to 5 years follow-up, HSCT was superior to ATZ in all the aforementioned aspects. Kaplan-Meier analysis at 5 years post treatment revealed superiority of HSCT in relapse rate (69.6 % vs 95.7 %, p = 0.027), MRI activity (54.5 % vs 75.1 %, p = 0.038) and disability worsening (57.1 % vs 90.9 %, p = 0.031). ATZ may halt disability progression early in the course of highly active RMS, but the disability starts accumulating later, while in HSCT patients disability improvement is consistent both 3 and 5 years after treatment onset.


Peroxisome proliferator-activated receptor-gamma agonists cause growth arrest and apoptosis in human ovarian carcinoma cell lines.
Peroxisome proliferator-activated receptor gamma (PPARgamma) is a member of the nuclear hormone receptor superfamily of ligand-activated transcription factors. PPARgamma agonists inhibit the growth of many types of cancers. To our knowledge, the effect of PPARgamma agonist on ovarian tumors is not reported. In this study, we used two human ovarian carcinoma cell lines (ES-2 and PA-1) to examine the effects of the PPARgamma agonists troglitazone (TGZ) and ciglitazone (CGZ) on cell survival. CGZ and TGZ inhibited viability in a dose-dependent manner in both types of ovarian cancer cells. The agonists also decreased cellular proliferation in association with an increase in the number of cells arrested in the G0/G1 phase of the cell cycle. Moreover, they increased apoptosis while increasing caspase-3 activity. Incubation of both the cell lines with the PPARgamma agonists led to upregulated PPARgamma expression. This effect appeared to be PPARgamma independent because the PPARgamma antagonist GW9662 did not reverse it. Along with the induction of apoptosis in ovarian cancer cells, protein expression levels of p53 and Bax markedly increased in response to the PPARgamma agonists. Our results demonstrated that PPARgamma agonists inhibited the viability of human ovarian cancer cells, at least partly by inducing apoptosis. As a result, these agonists may serve as future drugs for the prevention and treatment of ovarian cancer.


The role of host lymphoid populations in the response of mouse EMT6 tumor to photodynamic therapy.
Photodynamic therapy (PDT) treatment of murine EMT6 mammary sarcoma using Photofrin (10 mg/kg) and light (110 J/cm2) cured all these lesions growing in syngeneic BALB/c mice. In contrast, the same treatment produced initial ablation but no long-term cures of EMT6 tumors growing in either scid or nude mice, the immunodeficient strains sharing the same genetic background with BALB/c mice. No difference was detected in either the level of Photofrin accumulated per g of tumor tissue or the extent of tumor cell killing during the first 24 h after PDT of EMT6 tumors growing in BALB/c or scid mice. The assumption that the difference in tumor cures could be ascribed to the absence of functional lymphoid cells in scid and nude mice was supported by the results of experiments involving the adoptive T-cell transfer into scid mice or bone marrow transfer between BALB/c and scid mice. The adoptive transfer of splenic virgin T lymphocytes from BALB/c mice into scid mice performed 9 days before PDT of EMT6 tumors growing in the recipients was successful in delaying the recurrence of treated tumors. Adoptive transfer done immediately after PDT or 7 days after PDT had no obvious benefit. Even better improvement and a high cure rate of PDT-treated tumors was obtained with scid mice reconstituted with BALB/c bone marrow. In contrast, a marked drop in tumor cure rate was observed with BALB/c mice reconstituted with scid bone marrow. These results suggest that the activity of host lymphoid populations was essential for preventing the recurrence of EMT6 tumors following the PDT treatment used in this study. The contribution of PDT-induced immune reaction may, therefore, be of critical importance for the cure with at least some tumors.


Strength and power training did not modify cardiovascular responses to aerobic exercise in elderly subjects.
Resistance training increases muscle strength in older adults, decreasing the effort necessary for executing physical tasks, and reducing cardiovascular load during exercise. This hypothesis has been confirmed during strength-based activities, but not during aerobic-based activities. This study determined whether different resistance training regimens, strength training (ST, constant movement velocity) or power training (PT, concentric phase performed as fast as possible) can blunt the increase in cardiovascular load during an aerobic stimulus. Older adults (63.9 ± 0.7 years) were randomly allocated to: control (N = 11), ST (N = 13, twice a week, 70-90% 1-RM) and PT (N = 15, twice a week, 30-50% 1-RM) groups. Before and after 16 weeks, oxygen uptake (VO(2)), systolic blood pressure (SBP), heart rate (HR), and rate pressure product (RPP) were measured during a maximal treadmill test. Resting SBP and RPP were similarly reduced in all groups (combined data = -5.7 ± 1.2 and -5.0 ± 1.7%, respectively, P < 0.05). Maximal SBP, HR and RPP did not change. The increase in measured VO(2), HR and RPP for the increment in estimated VO(2) (absolute load) decreased similarly in all groups (combined data = -9.1 ± 2.6, -14.1 ± 3.9, -14.2 ± 3.0%, respectively, P < 0.05), while the increments in the cardiovascular variables for the increase in measured VO(2) did not change. In elderly subjects, ST and PT did not blunt submaximal or maximal HR, SBP and RPP increases during the maximal exercise test, showing that they did not reduce cardiovascular stress during aerobic tasks.


K(ICG) value, a reliable real-time estimator of graft function, accurately predicts outcomes in adult living-donor liver transplantation.
Reliable monitoring enabling evaluation of graft function is crucial after living-donor liver transplantation (LDLT). A method to identify poor graft function at an early postoperative period would allow opportune intensive clinical management to bring about further improvements in LDLT outcomes. This study assessed the reliability of the indocyanine green (ICG) elimination rate constant (K(ICG)) value as an estimator of graft function and determined the actual temporal changes of K(ICG) after LDLT. K(ICG) values were measured using a noninvasive method in 30 adult recipients up to 28 days after LDLT. The receptor index (LHL15) based on liver scintigraphy, and graft parenchymal damage score based on histopathological findings were evaluated after LDLT and correlated well with simultaneous K(ICG). Thus, K(ICG) measured by noninvasive method was confirmed as accurately evaluating graft function. Changes of K(ICG) after LDLT in recipients with good graft function were maintained, after some falls in the early periods, and had a significant difference compared with those for recipients without good graft function; moreover, there were already significant differences in K(ICG) 24 hours after LDLT. Mean transit time reflecting systemic hemodynamics revealed that recipients without good outcomes fell into an unstable systemic hemodynamic state, and effective hepatic blood flow has a large influence on liver regeneration after LDLT. In conclusion, we suggested that K(ICG) values can predict clinical outcomes at the early postoperative period after LDLT by sharply reflecting the influence of systemic dynamics on splanchnic circulation.


Does Fe(2+) in olivine-based interstellar grains play any role in the formation of H2? Atomistic insights from DFT periodic simulations.
Using periodic DFT-D2 methods, atomistic simulations of interstellar H adsorption and H2 formation on a (010) Fe-containing olivine surface are presented. At variance with the (010) Mg2SiO4 surface and key to these processes are the large Fe/H interaction energies, suggesting that olivine surfaces are good reservoirs of H atoms for subsequent recombination to form H2.


The serological differentiation of acute and chronic schistosomiasis japonica using IgA antibody to egg antigen.
Two groups of Schistosoma japonicum infected patients (acute and chronic) and non-infected individuals were studied using IgA antibody to egg antigen (SEA) and IgG and IgM antibodies to keyhole limpet haemocyanin (KLH). The means and standard deviation of the optical density in ELISA of acute, chronic and negative groups for IgA anti-SEA were 583 +/- 124.7, 98.2 +/- 78.8 and 82.2 +/- 39.3, respectively. There was a statistically significance between acute patients and chronic patients (P < 0.01). The means and standard deviation of IgG and IgM antibodies to KLH were 501.5 +/- 150.6, 113.0 +/- 79.1, 28.8 +/- 56.3 and 413.6 +/- 148.5, 70.2 +/- 14.8, 65.3 +/- 45.3, respectively. The detection results of IgA to SEA compared with the IgG and IgM to KLH did not demonstrate a significant difference (P < 0.01). The sensitivities of IgA to SEA and IgG and IgM antibodies to KLH for the detection of acute infection were 95.24%, 90.48% and 85.71%, respectively. Therefore, this study showed that the detection of IgA to SEA is also a useful new method for the serological differentiation of acute and chronic schistosomiasis japonica in humans.


Simultaneous multiple organ perfusion and transplantation.
This study demonstrates that multiple organ perfusion (kidney, heart, and pancreas) for 24 hours is a satisfactory method for transplantation. There was no evidence of perfusion damage by functional or histologic parameters. The potential clinical application of these findings is evident.


Thioaptamer decoy targeting of AP-1 proteins influences cytokine expression and the outcome of arenavirus infections.
Viral haemorrhagic fever (VHF) is caused by a number of viruses, including arenaviruses. The pathogenesis is believed to involve dysregulation of cytokine production. The arenaviruses Lassa virus and Pichinde virus have a tropism for macrophages and other reticuloendothelial cells and both appear to suppress the normal macrophage response to virus infection. A decoy thioaptamer, XBY-S2, was developed and was found to bind to AP-1 transcription factor proteins. The P388D1 macrophage-like cell line contains members of the AP-1 family which may act as negative regulators of AP-1-controlled transcription. XBY-S2 was found to bind to Fra-2 and JunB, and enhance the induction of cytokines IL-6, IL-8 and TNF-alpha, while reducing the binding to AP-1 promoter elements. Administration of XBY-S2 to Pichinde virus-infected guinea pigs resulted in a significant reduction in Pichinde virus-induced mortality and enhanced the expression of cytokines from primary guinea pig macrophages, which may contribute to its ability to increase survival of Pichinde virus-infected guinea pigs. These data demonstrate a proof of concept that thioaptamers can be used to modulate the outcome of in vivo viral infections by arenaviruses by the manipulation of transcription factors involved in the regulation of the immune response.


Changes in colonic mucosal permeability in mouse colitis induced with dextran sulfate sodium.
In this study we examined changes in colonic mucosal permeability induced by dextran sulfate sodium (DSS) during the acute phase of mouse colitis. To induce colitis, the mice were given drinking water containing 5% (w/v) DSS (MW = 40,000) ad libitum. Colonic mucosal permeability was evaluated by the permeation of Evans blue (EB) from the lumen into the wall of the colon on 1, 2, 3 and 7 days postadministration of DSS. Mucosal changes were also histologically examined daily for 7 days postadministration. The permeation of EB increased significantly by days 3 and 7 postadministration. Histological analysis showed that crypt loss was the initial change, with no inflammatory process and the surface mucosal epithelial cells remained morphologically intact. These histological changes developed on 2 to 3 days postadministration. Erosion was first recognized at 5 days postadministration. These findings indicated that the increase in colonic mucosal permeability may have occurred in 3 days postadministration, and the increase in mucosal permeability occurred before the appearance of the inflammatory process. This suggests that an increase in colonic mucosal permeability, leading to the destruction of mucosal barrier function, may play an important role in the induction of DSS-induced murine colitis.


Paraplegia after lumbosacral nerve root block: report of three cases.
Lumbar nerve root blocks and epidural steroid injections are frequently employed in the management of degenerative conditions of the lumbar spine, but relatively few papers have been published that address the complications associated with these interventions. Serious complications include epidural abscess, arachnoiditis, epidural hematoma, cerebrospinal fluid fistula and hypersensitivity reaction to injectate. Although transient paraparesis has been described after inadvertent intrathecal injection, an immediate and lasting deficit has not been previously described as sequelae of a nerve root block. We present three cases in which either persisting paraplegia or paraparesis occurred immediately after administration of a lumbar nerve root block and propose a mechanism for this devastating but previously unreported complication. Case reports of three patients. Three patients, two women and one man ranging in age from 42 to 64 years, underwent three procedures performed at three different facilities, in the hands of two different injectionists. In each instance, penetration of the dura was not thought to have occurred. In two procedures the needles were placed transforamenally, one at L3-4 on the left and one at L3-4 on the right, and in the third the needle tip was placed immediately lateral to the S1 nerve root. Patient follow-up data from medical office records. In each case, needle placement was verified with injection of a contrast media in conjunction with either computerized tomography or biplanar fluoroscopy. No backbleeding or cerebrospinal fluid was encountered upon aspiration in any of the procedures. Magnetic resonance imaging (MRI) was performed within 48 hours of injury in all patients. In each patient, paraplegia suddenly ensued after instillation of the steroid solution and, in each instance, postprocedure MRI revealed increased signal in the low thoracic spinal cord on T2-weighted imaging consistent with edema. The sudden onset of neurological deficit and the imaging changes noted in the spinal cord point to a vascular explanation for these injuries. We postulate that in these patients the spinal needle either penetrated or caused injury to an abnormally low dominant radiculomedullary artery, a recognized anatomical variant. This vessel, also known as the artery of Adamkiewicz, in 85% of individuals arises between T9 and L2, usually from the left, but in a minority of people may arise from the lower lumbar spine and rarely even from as low as S1. The artery of Adamkiewicz travels with the nerve root through the neural foramen and irrigates the anterior spinal artery. Injury of it or injection of particulate matter into it, as what may happen with the commonly used epidural steroid injectates, may result in infarction of the lower thoracic spinal cord, producing the clinical and imaging findings seen in these three patients. We present the cases of three patients who had lasting paraplegia or paraparesis after the performance of a nerve root block. We propose that the mechanism for this rare but devastating complication is the concurrence of two uncommon circumstances, the presence of an unusually low origin of the artery of Adamkiewicz and an undetected intraarterial penetration of the procedure needle.


Amyloid, Vascular, and Resilience Pathways Associated with Cognitive Aging.
To investigate the multifactorial processes underlying cognitive aging based on the hypothesis that multiple causal pathways and mechanisms (amyloid, vascular, and resilience) influence longitudinal cognitive decline in each individual through worsening brain health. We identified 1,230 elderly subjects (aged ≥50 years) with an average of 4.9 years of clinical follow-up and with amyloid positron emission tomography, diffusion tensor imaging, and structural magnetic resonance imaging scans from the population-based Mayo Clinic Study of Aging. We examined imaging markers of amyloid and brain health (white matter microstructural integrity and cortical thinning), systemic vascular health preceding the imaging markers, and early to midlife intellectual enrichment to predict longitudinal cognitive trajectories. We used latent growth curve models for modeling longitudinal cognitive decline. All the pathways (amyloid, vascular, resilience) converged through their effects on cortical thinning and worsening cognition and together explained patterns in cognitive decline. Resilience and vascular pathways (aging process, sex differences, education/occupation, and systemic vascular health) had significant impact on white matter microstructural integrity. Education/occupation levels contributed to white matter integrity through systemic vascular health. Worsening white matter integrity contributed to significant cortical thinning and subsequently longitudinal cognitive decline. Baseline amyloidosis contributed to a significant proportion of cognitive decline that accelerated with longer follow-up times, and its primary impact was through cortical thinning. We developed an integrated framework to help explain the dynamic and complex process of cognitive aging by considering key causal pathways. Such an approach is important for both better comprehension of cognitive aging processes and will aid in the development of successful intervention strategies. ANN NEUROL 2019;86:866-877.


A standardized care plan is associated with shorter hospital length of stay in patients undergoing pancreaticoduodenectomy.
In this retrospective review, we evaluate a standardized care plan (SCP) for patients undergoing pancreaticoduodenectomy, which included selective placement of feeding jejunostomy tubes (FJTs) and a perioperative fast-track recovery pathway (FTRP). A review of 242 patients undergoing pancreaticoduodenectomy was completed. Patients treated pre- and post-SCP implementation were compared. Univariate comparison followed by multivariable linear regression were performed to identify predictors of hospital length of stay (HLOS). SCP patients (n = 100) were slightly older but otherwise similar to pre-SCP patients (n = 142). FJT placement occurred less frequently in SCP patients (38 versus 94%, P < 0.001). All SCP patients were initiated on the FTRP. Among SCP patients, an oral diet was introduced earlier (5 versus 8.5 d, P < 0.001) and HLOS was shorter (11 versus 13 d, P = 0.015). Readmission rates were similar. Following adjustment with linear regression, we confirmed SCP status as a predictor of HLOS. To assess SCP components, HLOS was evaluated separately based on FTRP status and FJT placement. Although both were highly associated with HLOS, neither was independently predictive in multivariable analysis. Implementation of an SCP resulted in shorter HLOS without an increase in readmissions. Future studies are necessary to identify specific components of SCPs that most influence outcomes.


Laryngeal afferent inputs to the nucleus of the solitary tract.
The following study was undertaken to examine the integration of laryngeal afferent inputs within the nucleus of the solitary tract (NTS), the primary site of termination of laryngeal afferent fibers. Intracellular recordings were obtained from 63 cells that responded to electrical stimulation of the superior laryngeal nerve (SLN) with an excitatory postsynaptic potential (EPSP; n = 49), an excitatory-inhibitory postsynaptic potential (EPSP-IPSP) sequence (n = 13), or an IPSP (n = 1). Mechanical stimulation of laryngeal mechanoreceptors revealed a variety of response patterns (e.g., slowly and rapidly adapting depolarizations or hyperpolarizations). Two types of response to increasing SLN stimulus frequency were observed. In 11 cells SLN-evoked EPSP amplitude at 10 Hz was only 47 +/- 4% of the amplitude at 1 Hz, while in 6 cells EPSP amplitude at 10 Hz was virtually identical (93 +/- 3%) to that at 1 Hz. Time-dependent inhibitory interactions occurred between SLN inputs to NTS neurons at intervals between 50 and 400 ms and in the absence of any change in membrane potential. NTS neuronal responses to brief activation of laryngeal mechanoreceptors correspond well to discharge patterns described for individual laryngeal mechanoreceptors. Frequency-dependent filtering and time-dependent inhibitory interactions might modify NTS neuronal responses during more intense stimulation of laryngeal afferents.


Objectivity and ethics in environmental health science.
During the past several decades, philosophers of science and scientists themselves have become increasingly aware of the complex ways in which scientific knowledge is shaped by its social context. This awareness has called into question traditional notions of objectivity. Working scientists need an understanding of their own practice that avoids the naïve myth that science can become objective by avoiding social influences as well as the reductionist view that its content is determined simply by economic interests. A nuanced perspective on this process can improve research ethics and increase the capacity of science to contribute to equitable public policy, especially in areas such as environmental and occupational health, which have direct implications for profits, regulation, legal responsibility, and social justice. I discuss research into health effects of the 1979 accident at Three Mile Island near Harrisburg, Pennsylvania, USA, as an example of how scientific explanations are shaped by social concepts, norms, and preconceptions. I describe how a scientific practice that developed under the influence of medical and nuclear physics interacted with observations made by exposed community members to affect research questions, the interpretation of evidence, inferences about biological mechanisms in disease causation, and the use of evidence in litigation. By considering the history and philosophy of their disciplines, practicing researchers can increase the rigor, objectivity, and social responsibility of environmental health science.


H-ras expression in immortalized keratinocytes produces an invasive epithelium in cultured skin equivalents.
Ras proteins affect both proliferation and expression of collagen-degrading enzymes, two important processes in cancer progression. Normal skin architecture is dependent both on the coordinated proliferation and stratification of keratinocytes, as well as the maintenance of a collagen-rich basement membrane. In the present studies we sought to determine whether expression of H-ras in skin keratinocytes would affect these parameters during the establishment and maintenance of an in vitro skin equivalent. Previously described cdk4 and hTERT immortalized foreskin keratinocytes were engineered to express ectopically introduced H-ras. Skin equivalents, composed of normal fibroblast-contracted collagen gels overlaid with keratinocytes (immortal or immortal expressing H-ras), were prepared and incubated for 3 weeks. Harvested tissues were processed and sectioned for histology and antibody staining. Antigens specific to differentiation (involucrin, keratin-14, p63), basement-membrane formation (collagen IV, laminin-5), and epithelial to mesenchymal transition (EMT; e-cadherin, vimentin) were studied. Results showed that H-ras keratinocytes produced an invasive, disorganized epithelium most apparent in the lower strata while immortalized keratinocytes fully stratified without invasive properties. The superficial strata retained morphologically normal characteristics. Vimentin and p63 co-localization increased with H-ras overexpression, similar to basal wound-healing keratinocytes. In contrast, the cdk4 and hTERT immortalized keratinocytes differentiated similarly to normal unimmortalized keratinocytes. The use of isogenic derivatives of stable immortalized keratinocytes with specified genetic alterations may be helpful in developing more robust in vitro models of cancer progression.


Identify sequence of events likely to result in severe crash outcomes.
The current practice of crash characterization in highway engineering reduces multiple dimensions of crash contributing factors and their relative sequential connections, crash sequences, into broad definitions, resulting in crash categories such as head-on, sideswipe, rear-end, angle, and fixed-object. As a result, crashes that are classified in the same category may contain many different crash sequences. This makes it difficult to develop effective countermeasures because these crash categorizations are based on the outcomes rather than the preceding events. Consequently, the efficacy of a countermeasure designed for a specific type of crash may not be appropriate due to different pre-crash sequences. This research seeks to explore the use of event sequence to characterize crashes. Additionally, this research seeks to identify crash sequences that are likely to result in severe crash outcomes so that researchers can develop effective countermeasures to reduce severe crashes. This study utilizes the sequence of events from roadway departure crashes in the Fatality Analysis Reporting System (FARS), and converts the information to form a new categorization called "crash sequences." The similarity distance between each pair of crash sequences were calculated using the Optimal Matching approach. Cluster analysis was applied to group crash sequences that are etiologically similar in terms of the similarity distance. A hybrid model was constructed to mitigate the potential sample selection bias of FARS data, which is biased toward more severe crashes. The major findings include: (1) in terms of a roadway departure crash, the crash sequences that are most likely to result in high crash severity include a vehicle that first crosses the median or centerline, runs-off-road on the left, and then collides with a roadside fixed-object; (2) seat-belt and airbag usage reduces the probability of dying in a roadway departure crash by 90%; and (3) occupants who are seated on the side of the vehicle that experience a direct impact are 2.6 times more likely to die in a roadway departure crash than those not seated on the same side of the vehicle where the impact occurs.


Transsphenoidal computer-navigation-assisted deflation of a balloon after endovascular occlusion of a direct carotid cavernous sinus fistula.
A 49-year-old woman with a direct posttraumatic carotid cavernous fistula (CCF) was treated with detachable balloons via a transcarotid route. After the procedure, her intracranial bruit, conjunctival injection, and orbital congestion were cured, but the preexistent sixth nerve palsy deteriorated. CT showed one balloon positioned in the posterior portion of the right cavernous sinus and was regarded to be responsible for nerve compression. After surgical exposure by use of a transnasal-transsphenoidal approach under 3D navigation control, this balloon was deflated by puncture with a 22-gauge needle. The previously described symptoms resolved after balloon deflation. This report presents a rare complication of endovascular treatment of direct CCF and a new microsurgical approach to a balloon in a case of nerve compression.