Electronic cigarette exposure disrupts airway epithelial barrier function and exacerbates viral infection.
The use of electronic cigarettes (e-cigs), especially among teenagers, has reached alarming and epidemic levels, posing a significant threat to public health. However, the short- and long-term effects of vaping on the airway epithelial barrier are unclear. Airway epithelial cells are the forefront protectors from viruses and pathogens. They contain apical junctional complexes (AJCs), which include tight junctions (TJs) and adherens junctions (AJs) formed between adjacent cells. Previously, we reported respiratory syncytial virus (RSV) infection, the leading cause of acute lower respiratory infection-related hospitalization in children and high-risk adults, induces a "leaky airway" by disrupting the epithelial AJC structure and function. We hypothesized chemical components of e-cigs disrupt airway epithelial barrier and exacerbate RSV-induced airway barrier dysfunction. Using confluent human bronchial epithelial (16HBE) cells and well-differentiated normal human bronchial epithelial (NHBE) cells, we found that exposure to extract and aerosol e-cig nicotine caused a significant decrease in transepithelial electrical resistance (TEER) and the structure of the AJC even at noncytotoxic concentrations. Western blot analysis of 16HBE cells exposed to e-cig nicotine extract did not reveal significant changes in AJC proteins. Exposure to aerosolized e-cig cinnamon or menthol flavors also induced barrier disruption and aggravated nicotine-induced airway barrier dysfunction. Moreover, preexposure to nicotine aerosol increased RSV infection and the severity of RSV-induced airway barrier disruption. Our findings demonstrate that e-cig exposure disrupts the airway epithelial barrier and exacerbates RSV-induced damage. Knowledge gained from this study will provide awareness of adverse e-cig respiratory effects and positively impact the mitigation of e-cig epidemic.NEW & NOTEWORTHY Electronic cigarette (e-cig) use, especially in teens, is alarming and at epidemic proportions, threatening public health. Our study shows that e-cig nicotine exposure disrupts airway epithelial tight junctions and increases RSV-induced barrier dysfunction. Furthermore, exposure to aerosolized flavors exaggerates e-cig nicotine-induced airway barrier dysfunction. Our study confirms that individual and combined components of e-cigs deleteriously impact the airway barrier and that e-cig exposure increases susceptibility to viral infection.


Neurosyphilis.
Neurosyphilis is caused by the bacterium Treponema pallidum subspecies pallidum (T. pallidum). The organism gains entry into the central nervous system (CNS) early (primary syphilis or chancre phase) in the course of infection. While most patients are able to mount an immune response that effectively clears CNS invasion without long-term complications, a minority go on to develop asymptomatic or symptomatic neurosyphilis. Neurosyphilis has been divided into early and late stages. The early stages include asymptomatic meningitis, symptomatic meningitis, gumma, and meningovascular syphilis, while the late stages include dementia paralytica and tabes dorsalis. Ocular and otologic syphilis can occur at any time but often accompany the acute meningitis of early neurosyphilis. The diagnosis of symptomatic neurosyphilis requires meeting clinical, serologic, and cerebrospinal fluid (CSF) criteria, while the diagnosis of asymptomatic neurosyphilis relies on serologic and CSF criteria alone. In the last several decades, a persistent rise in syphilitic meningitis and other forms of early neurosyphilis have been seen in the human immunodeficiency virus-positive population, principally in men who have sex with men. This article reviews the clinical presentation, diagnosis, and treatment of neurosyphilis, and it addresses the controversy regarding the role of lumbar puncture early in the course of infection.


Responsiveness of plasma 2- and 4-hydroxycatecholestrogens to training and to graduate submaximal and maximal exercise in an untrained woman. Interuniversity Project on Reproductive Endocrinology in Women and Exercise.
A single-subject experimental design was used to obtain some preliminary findings on the plasma responses of catecholestrogens (CE) to acute exercise and brief, but exhaustive training on a cycle ergometer. One previously untrained eumenorrheic female (body fat: 26% VO2max: 43.3 ml x kg(-1) x min[-1]) participated in this study. Resting CE levels were for "total" (unconjugated + conjugated) 2-hydroxyestrogens (2-OHE) 162pg/ ml and 350 pg/ml in the follicular (FPh) and luteal phase (LPh), respectively. Plasma total 4-hydroxyestrogen (4-OHE) levels were 41 pg/ml in the FPh and 66 pg/ml in the LPh. For "total" 2-methoxyestrogens (2-MeOE), we found 257 pgl/ml in the FPh and 374 pg/ml in the LPh. Resting levels of 2-hydroxy CE following a period of brief, intensive training were decreased during the LPh (2-OHE: -38%; 2-MeOE: -19%), whereas 4-hydroxy CE were unaffected. After training, the formation of CE as expressed by the 2-OHE:E and 4-OHE:E ratios, was increased by 75% and 200% at rest, respectively. CE activity or O-methylation, as estimated from the 2-MeOE:2-OHE ratio, was higher following training (FPh: +22%; LPh: +30%). During acute exercise before training, we observed a small rise proportional to the exercise intensity in the plasma "total" primary estrogen concentrations (FPh: +28%; LPh: +16%), and no changes in either 2-OHE or 2 MeOE levels. Plasma concentrations of 4-OHE, however, doubled during maximal exercise intensity. The 2-OHE:E and 2-MeOE:2-OHE ratios did not alter during incremental exercise. Training effects on acute exercise responses were only noticed for 4-OHE, which contrary to pre-training conditions, now progressively decreased. The major findings of this study are that in response to training: a) during rest, a greater proportion of CE are formed from a lower amount of precursor hormone, b) the rate of O-methylation of CE increases.


The effect of age on the development of hypercholesterolemia in rabbits fed semipurified diets containing casein.
Young and adult male rabbits were alternately fed semipurified diets and a commercial diet over a period of 57 weeks. The semipurified diets, containing either casein or soy protein, and the commercial diet were fed either ad libitum or on a restricted basis. When a restricted feeding regime was applied, both in the young and adult rabbits, significantly higher levels of serum cholesterol were observed in the animals fed casein compared with their counterparts fed soy protein. However, during the first period of feeding the semipurified diets, the hypercholesterolemic response of the casein diet was significantly greater in the young than in the adult rabbits. During the second and third period, no significant differences in cholesterolemic response were observed between young and adult rabbits fed casein diets. Further, the cholesterolemic response to semipurified diets containing casein was progressively lower during the second and third period. Similar results were found when the rabbits were fed ad libitum. However, in the adult rabbits no significant differences were observed between the rabbits fed casein and soy protein. Thus, the results of this study show that adult rabbits are less susceptible to the induction of hypercholesterolemia by feeding casein diets than are young ones.


AUDITORY NERVE: ELECTRICAL STIMULATION IN MAN.
Auditory perceptions produced in a person deaf to acoustic stimulation were studied by electrically exciting the auditory nerve through permanently implanted electrodes. Pulsed current as small as 1 microampere peak-to-peak could be perceived. Pitch, as reported by the subject, varied with electrode selection, current amplitude, and pulse repetition rate from about 70 to at least 300 pulses per second. Loudness increased with amplitude and duration of pulse stimuli, and to a lesser extent with repetition rate. The total range in amplitude of the stimulus, from threshold to an uncomfortable loudness, was 15 to 20 decibels. Simultaneous stimulation in separate electrodes produced a number of complex effects.


PI3K/AKT pathway mutations cause a spectrum of brain malformations from megalencephaly to focal cortical dysplasia.
Malformations of cortical development containing dysplastic neuronal and glial elements, including hemimegalencephaly and focal cortical dysplasia, are common causes of intractable paediatric epilepsy. In this study we performed multiplex targeted sequencing of 10 genes in the PI3K/AKT pathway on brain tissue from 33 children who underwent surgical resection of dysplastic cortex for the treatment of intractable epilepsy. Sequencing results were correlated with clinical, imaging, pathological and immunohistological phenotypes. We identified mosaic activating mutations in PIK3CA and AKT3 in this cohort, including cancer-associated hotspot PIK3CA mutations in dysplastic megalencephaly, hemimegalencephaly, and focal cortical dysplasia type IIa. In addition, a germline PTEN mutation was identified in a male with hemimegalencephaly but no peripheral manifestations of the PTEN hamartoma tumour syndrome. A spectrum of clinical, imaging and pathological abnormalities was found in this cohort. While patients with more severe brain imaging abnormalities and systemic manifestations were more likely to have detected mutations, routine histopathological studies did not predict mutation status. In addition, elevated levels of phosphorylated S6 ribosomal protein were identified in both neurons and astrocytes of all hemimegalencephaly and focal cortical dysplasia type II specimens, regardless of the presence or absence of detected PI3K/AKT pathway mutations. In contrast, expression patterns of the T308 and S473 phosphorylated forms of AKT and in vitro AKT kinase activities discriminated between mutation-positive dysplasia cortex, mutation-negative dysplasia cortex, and non-dysplasia epilepsy cortex. Our findings identify PI3K/AKT pathway mutations as an important cause of epileptogenic brain malformations and establish megalencephaly, hemimegalencephaly, and focal cortical dysplasia as part of a single pathogenic spectrum.


Three cases of intravenous sodium benzoate and sodium phenylacetate toxicity occurring in the treatment of acute hyperammonaemia.
Intravenous sodium benzoate and sodium phenylacetate have been used successfully in the treatment of acute hyperammonaemia in patients with urea cycle disorders. They provide alternative pathways for waste nitrogen disposal and help maintain nitrogen homeostasis. However, we report three patients with hyperammonaemia who received inappropriate doses of intravenous sodium benzoate and sodium phenylacetate that resulted in severe complications. Ambiguous medical prescriptions and inadequate cross-checking of drug dosage by physicians, nurses and pharmacists were the main causes of these incidents. All the patients presented with alteration in mental status, Kussmaul respiration and a partially compensated metabolic acidosis with an increased anion gap. Two patients developed cerebral oedema and hypotension and died. The third survived after haemodialysis. Plasma levels of benzoate and phenylacetate were excessively high. The possible mechanisms of toxicity, management and safety measures are discussed.


Arginylation-dependent regulation of a proteolytic product of talin is essential for cell-cell adhesion.
Talin is a large scaffolding molecule that plays a major role in integrin-dependent cell-matrix adhesion. A role for talin in cell-cell attachment through cadherin has never been demonstrated, however. Here, we identify a novel calpain-dependent proteolytic cleavage of talin that results in the release of a 70-kD C-terminal fragment, which serves as a substrate of posttranslational arginylation. The intracellular levels of this fragment closely correlated with the formation of cell-cell adhesions, and this fragment localized to cadherin-containing cell-cell contacts. Moreover, reintroduction of this fragment rescued the cell-cell adhesion defects in arginyltransferase (Ate1) knockout cells, which normally have a very low level of this fragment. Arginylation of this fragment further enhanced its ability to rescue cell-cell adhesion formation. In addition, arginylation facilitated its turnover, suggesting a dual role of arginylation in its intracellular regulation. Thus, our work identifies a novel proteolytic product of talin that is regulated by arginylation and a new role of talin in cadherin-dependent cell-cell adhesion.


Empowering Native Adolescents: Responsibility for Their Health Behaviors.
Objectives: In this study, we assess the impact of a home-based diabetes prevention program, Together on Diabetes (TOD), on adolescent responsibility-taking for tasks related to diabetes risk. Methods: Participants were Native American youth ages 10-19 with or at risk of type 2 diabetes who participated in a 12-session, 6-month diabetes prevention program with an adult caretaker. Assessments completed at baseline, 6-month, and 12-month follow-up include demographics and the Diabetes and Obesity Task Sharing (DOTS) Questionnaire. We used latent class analysis (LCA) at baseline to examine heterogeneity in DOTS responses. We identified 3 classes (adolescent, shared, caretaker). We used latent transition analysis to examine stability and change in latent status at baseline, 6- and 12-month follow-up. Results: At baseline, the mean age of participants was 13.6 years and 55.9% were boys. From baseline to 6-month follow-up, the adolescent class was most stable, whereas the shared and caretaker classes were less stable. For participants who transition from the adolescent class, most transition to shared class compared to caretaker class. Conclusions: TOD helps to empower Native American adolescents to take responsibility for their health and engage with their caregivers in these decisions.


Two-Needle Technique for Lumbar Radiofrequency Medial Branch Denervation: A Technical Note.
Radiofrequency ablation (RFA) of the medial branches of the dorsal rami has been reported to relieve facet joint-related back pain for 6 months to 1 year in 60% of patients. Although providing benefit in a significant proportion of patients, there remains a group of patients who do not experience any pain relief from RFA or experience only benefit from the ablation for a short period. Failure of RFA has been attributed to technical failure of coagulating the nerve or coagulation of a minimal section of the nerve, allowing for early reinnervation. Increasing the success rate and duration of relief may require techniques that increase the likelihood of successful nerve ablation over a relevant distance by maximizing lesion size. The aim of this technical note is to detail a modification to the current commonly used lumbar medial branch radiofrequency (RF) denervation approach to increase lesion size. This is a technical report describing a novel two-needle approach to lumbar RF medial branch denervation. Large private interventional pain management institute. A dual needle placement of two 10-mm active tip RF cannulas separated by 6 mm is used to optimally contact the superior articular process (SAP) from its ventral to dorsal borders, which encompasses the anticipated course of the medial branch nerves. The described technique creates a lesion that we estimate to be 11.0-mm wide and 11.6-mm long along the course of the medial branch adjacent to the SAP ensuring adequate coverage and treatment. This report does not encompass a systematic evaluation of the clinical safety and efficacy of the two-needle RFA approach. Future studies will have to assess the long-term efficacy and safety of the approach. The detailed two-needle approach to lumbar RF medial branch denervation appears to be promising in terms of projected treatment success by coagulating a large volume of tissue, in a cost- and time-efficient manner.


The influence of cyclosporin A on proteininduced uveitis in the rabbit.
Until now immunosuppressive drugs have mainly been used to treat ocular diseases considered to have an autoimmune pathogenesis. The authors investigated whether cyclosporin A (CsA) could also prevent intraocular inflammation mediated by a foreign antigen. To this purpose, uveitis was induced by injection of human serum albumin (HSA) into the vitreous of rabbits. Subcutaneous injection of CsA prevented the induction of uveitis. Treatment of CsA had to be started at the time of intravitreal antigen injection and did not suppress the reaction when started at the onset of uveitis. Suppression of uveitis correlated with an inhibition of the antibody response against the injected HSA. Animals in which uveitis was suppressed by CsA did not develop a recurrent uveitis after intravenous challenge with the antigen, but did develop a primary inflammatory response after a repeated injection of HSA into the vitreous. The most likely interpretation of the findings presented in this paper are as follows. CsA blocks T helper cells through an inhibition of IL-2 gene activation. This in turn blocks release of other T helper cell cytokines which are essential for the activation of B lymphocytes into antibody producing plasma cells. These observations thus show that CsA can suppress both cell-mediated as well as antibody-mediated models of uveitis.


Functional outcome of displaced radial head fractures in children treated by elastic stable intramedullary nailing.
The objectives of this study were to retrospectively investigate the clinical and radiological outcomes of displaced radial head fractures in children treated by elastic stable intramedullary nailing (ESIN) and evaluate the functional outcome of these injuries using the short version of the Disabilities of the Arm, Shoulder and Hand outcome questionnaire (Quick DASH). A total of 24 patients (nine males and 15 females) fulfilled the inclusion criteria. The mean age at the time of injury was 10.7±2.8 years (range: 9.5-16.33 years). Before surgery, the mean angulation was 53.8°±18.4° (range: 28°-82°) differentiating fractures on the basis of Judet's classification. The mean Quick DASH score was a good 4 (range: 0-15.9). Functional outcomes were similar irrespective of the severity of fracture displacement (P>0.05), presence or absence of associated fracture (P>0.05), and time immobilized (P>0.05). The Quick DASH score was better in children younger than 9 years of age (1.62) than children older than 9 years of age (4.95), but without a statistically significant difference (P=0.058). Children with displaced radial head fractures treated with ESIN showed good functional outcomes. Associated fracture injuries were not a predictive factor of functional outcome. Open reduction must be avoided as it carries an increased risk of complications.


Active and sedentary bouts in people after stroke and healthy controls: An observational study.
Understanding how both active and sedentary time is accumulated in people after stroke may help to better target interventions to reduce stroke recurrence. This study aimed to determine the difference between stroke and healthy controls in (a) time spent in sedentary and active behaviour, (b) frequency of short and long active and sedentary bouts and (c) time spent in short and long active and sedentary bouts. Analysis of secondary outcomes from a cross-sectional study. Participants were 42 community-dwelling people after stroke and 21 age-matched healthy controls. An activity monitor was used to collect free-living active and sedentary behaviour. Total active (standing and walking) and sedentary (lying, reclining and sitting) time was calculated in minutes per day. Bouts were categorized as short (<5 min, 5-15 min, 15-30 min) or long (>30 min). The frequency of and time spent in each bout were calculated. Relative to wear time, the stroke group spent 10% (95% confidence interval [CI] 3 to 17) more time in sedentary behaviour and had fewer long active bouts than the healthy controls. The stroke group spent 7% (95% CI 1-13) less time in long active bouts and 11% (95% CI 2-20) more time in long sedentary bouts than the healthy controls. Community-dwelling people after stroke spent less time in active behaviour and accumulated more sedentary time in bouts longer than 30 min compared with healthy controls. Increasing active time and breaking up long sedentary time warrants investigation in people after stroke.


Early signs of anomalous neural functional connectivity in healthy offspring of parents with bipolar disorder.
Bipolar disorder (BD) has been associated with dysfunctional brain connectivity and with family chaos. It is not known whether aberrant connectivity occurs before illness onset, representing vulnerability for developing BD amidst family chaos. We used resting-state functional magnetic resonance imaging (fMRI) to examine neural network dysfunction in healthy offspring living with parents with BD and healthy comparison youth. Using two complementary methodologies [data-driven independent component analysis (ICA) and hypothesis-driven region-of-interest (ROI)-based intrinsic connectivity], we examined resting-state fMRI data in 8-17-year-old healthy offspring of a parent with BD (n = 24; high risk) and age-matched healthy youth without any personal or family psychopathology (n = 25; low risk). ICA revealed that, relative to low-risk youth, high-risk youth showed increased connectivity in the ventrolateral prefrontal cortex (VLPFC) subregion of the left executive control network (ECN), which includes frontoparietal regions important for emotion regulation. ROI-based analyses revealed that high-risk versus low-risk youth had decreased connectivities between the left amygdala and pregenual cingulate, between the subgenual cingulate and supplementary motor cortex, and between the left VLPFC and left caudate. High-risk youth showed stronger connections in the VLPFC with age and higher functioning, which may be neuroprotective, and weaker connections between the left VLPFC and caudate with more family chaos, suggesting an environmental influence on frontostriatal connectivity. Healthy offspring of parents with BD show atypical patterns of prefrontal and subcortical intrinsic connectivity that may be early markers of resilience to or vulnerability for developing BD. Longitudinal studies are needed to determine whether these patterns predict outcomes.


Pharmacokinetic Interactions of Clinical Interest Between Direct Oral Anticoagulants and Antiepileptic Drugs.
Direct oral anticoagulants (DOACs), namely apixaban, dabigatran, edoxaban, and rivaroxaban are being increasingly prescribed among the general population, as they are considered to be associated to lower bleeding risk than classical anticoagulants, and do not require coagulation monitoring. Likewise, DOACs are increasingly concomitantly prescribed in patients with epilepsy taking, therefore, antiepileptic drugs (AEDs), above all among the elderly. As a result, potential interactions may cause an increased risk of DOAC-related bleeding or a reduced antithrombotic efficacy. The objective of the present review is to describe the pharmacokinetic interactions between AEDs and DOACs of clinical relevance. We observed that there are only few clinical reports in which such interactions have been described in patients. More data are available on the pharmacokinetics of both drugs classes which allow speculating on their potential interactions. Older AEDs, acting on cytochrome P450 isoenzymes, and especially on CYP3A4, such as phenobarbital, phenytoin, and carbamazepine are more likely to significantly reduce the anticoagulant effect of DOACs (especially rivaroxaban, apixaban, and edoxaban). Newer AEDs not affecting significantly CYP or P-gp, such as lamotrigine, or pregabalin are not likely to affect DOACs efficacy. Zonisamide and lacosamide, which do not affect significantly CYP activity in vitro, might have a quite safe profile, even though their effects on P-gp are not well-known, yet. Levetiracetam exerts only a potential effect on P-gp activity, and thus it might be safe, as well. In conclusion, there are only few case reports and limited evidence on interactions between DOACs and AEDs in patients. However, the overall evidence suggests that the interaction between these drug classes might be of high clinical relevance and therefore further studies in larger patients' cohorts are warranted for the future in order to better clarify their pharmacokinetic and define the most appropriate clinical behavior.


[Issues concerning the measurement and assessment of noise in workplaces].
The measurement and assessment of noise in workplaces are affected by several critical aspects. Only the equivalent sound pressure levels are measured and all the other acoustical parameters are almost always neglected; the non-acoustic information that characterize each measurement are not usually reported. The survey about noise in workplaces is not suited to be described by technical standards, but rather by flexible guidelines. The reference standard UNI EN ISO 9612/2011 does not highlight the contribution given by each work phase to the overall exposure level; the uncertainty evaluation may lead to unreliable data.


Aging, eye movements, and object-location memory.
This study investigated whether "intentional" instructions could improve older adults' object memory and object-location memory about a scene by promoting object-oriented viewing. Eye movements of younger and older adults were recorded while they viewed a photograph depicting 12 household objects in a cubicle with or without the knowledge that memory about these objects and their locations would be tested (intentional vs. incidental encoding). After viewing, participants completed recognition and relocation tasks. Both instructions and age affected viewing behaviors and memory. Relative to incidental instructions, intentional instructions resulted in more accurate memory about object identity and object-location binding, but did not affect memory accuracy about overall positional configuration. More importantly, older adults exhibited more object-oriented viewing in the intentional than incidental condition, supporting the environmental support hypothesis.


Plasma symmetric dimethylarginine reference limits from the Framingham offspring cohort.
Symmetric dimethylarginine (SDMA) is a by-product of protein methylation. Once released from proteins, SDMA is eliminated by the kidneys; consequently, plasma concentration has been suggested as a sensitive marker of renal function. Furthermore, recent work implicates SDMA in the pathogenesis of cardiovascular disease. To date, reference limits for SDMA plasma concentrations in healthy individuals are lacking. This study defined reference limits for plasma SDMA concentrations in 840 relatively healthy individuals of the Offspring Cohort from Framingham Heart Study (mean age 56 years, 61% women). Plasma SDMA concentrations were determined by LC-MS/MS using a stable isotope dilution assay. The median SDMA concentration in the reference sample was 0.37 μmol/L (Q1, Q3:0.32, 0.43 μmol/L) and the reference limits were 0.225 and 0.533 (2.5th and 97.5th percentile). In a multivariable regression model, serum creatinine, age and total homocysteine were positively associated with SDMA (p<0.001 for all), whereas the body mass index and diastolic blood pressure were inversely related to SDMA (p-values<0.01 and 0.03, respectively). This study reports plasma SDMA reference limits from the community-based Framingham Heart Study. Plasma SDMA concentration was related positively to advancing age, but inversely to renal function. These reference limits may allow the identification of individuals with raised plasma SDMA concentrations.


[The comparative morphology of Trichinella spiralis and Trichinella pseudospiralis].
The comparative study of the morphology of invasive larvae and mature males and females T. spiralis and T. pseudospiralis at the head end of the parasites has revealed 14 papillae which are located in 3 circles and 2 which are placed bilaterally, 2 amphidae, and a lanceolar stiletto with a spicular cutting edge. There are no differences in the structure of the heads of these two Trichinella species. The larval cuticle is decorated only with cross folds while longitudinal small-sized folds which are arranged in regular rows appeared in mature species. The vulva in females is located at the levels of a fifth of the body from the head, it is slit-like and oval, that in T. pseudospiralis is situated at the cuticular projection. The anus in T. pseudospiralis larvae is as a cross oval slit, that in adult females is as a hemisphere, and in adult males it is coincident with the copulation pore. As for the structure of the pseudobursa, differences are found in the size and shape of copulation processes, in the arrangement of pre- and postanal papillae against the cloaca, in the size of the formed cuticular projections in T. spiralis, in the shape of the cloacal hole. In the unscrewed copulatory bell, it opens transversely in T. spiralis and vertically in T. pseudospiralis.


Single-Cell Profiling of Premature Neonate Airways Reveals a Continuum of Myeloid Differentiation.
Single-cell genomic technologies hold great potential to advance our understanding of lung development and disease. A major limitation lies in accessing intact cells from primary lung tissues for profiling human airway health. Sampling methods such as endotracheal aspiration that are compatible with clinical interventions could enable longitudinal studies, the enrollment of large cohorts, and the development of novel diagnostics. To explore single-cell RNA sequencing profiling of the cell types present at birth in the airway lumen of extremely premature neonates (<28 wk gestation), we isolated cells from endotracheal aspirates collected from intubated neonates within the first hour after birth. We generated data on 10 subjects, providing a rich view of airway luminal biology at a critical developmental period. Our results show that cells present in the airways of premature neonates primarily represent a continuum of myeloid differentiation, including fetal monocytes (25% of total), intermediate myeloid populations (48%), and macrophages (2.6%). Applying trajectory analysis to the myeloid populations, we identified two trajectories consistent with the developmental stages of interstitial and alveolar macrophages, as well as a third trajectory presenting an alternative pathway bridging the distinct macrophage precursors. The three trajectories share many dynamic genes (N = 5,451), but also have distinct transcriptional changes (259 alveolar-specific, 666 interstitial-specific, and 285 bridging-specific). Overall, our results define cells isolated within the so-called "golden hour of birth" in extremely premature neonate airways, representing complex lung biology, and can be used in studies of human development and disease.


MEMS scanning micromirror for optical coherence tomography.
This paper describes an endoscopic-inspired imaging system employing a micro-electromechanical system (MEMS) micromirror scanner to achieve beam scanning for optical coherence tomography (OCT) imaging. Miniaturization of a scanning mirror using MEMS technology can allow a fully functional imaging probe to be contained in a package sufficiently small for utilization in a working channel of a standard gastroesophageal endoscope. This work employs advanced image processing techniques to enhance the images acquired using the MEMS scanner to correct non-idealities in mirror performance. The experimental results demonstrate the effectiveness of the proposed technique.


[Sisomycin pharmacokinetics in the perilymph and blood serum--an approach to predicting its ototoxic effect].
To elucidate the possibility of predicting the level of aminoglycoside antibiotic penetration into the fluids of the internal ear by the antibiotic blood levels, the pharmacokinetics of sisomicin in the perilymph and blood serum was studied on guinea pigs. The antibiotic was administered to the animals subcutaneously in doses of 50, 100 and 200 mg/kg. On the basis of the comparison of the sisomicin concentrations in the perilymph normalized against the dose it was concluded that the pharmacokinetics of sisomicin in the perilymph and blood serum of the animals was linear. Comparison of the areas under the curves of the antibiotic concentration versus time in the perilymph (AUCp) and blood serum (AUCs) showed that the tissue availability of the antibiotic in this study characterized by its penetration into the perilymph and defined by the ratio of the AUCp to AUCs amounted to 55 per cent. In a two-compartment model it was not possible to predict the antibiotic levels in the perilymph by concentrations in the blood. However, by the antibiotic blood levels it was possible to characterize in a complex the pharmacokinetic behaviour of the antibiotic in the perilymph by predicting the areas under the respective curves of the antibiotic concentration versus time. The proportional relation between the values of the AUCp and AUCs suggested that the level of the antibiotic penetration into the internal ear and consequently the intensity of the potential ototoxic effect could be more reliably predicted not by separate values of the antibiotic concentration but by the areas under curves of aminoglycoside concentrations versus time.


Current perspectives in drug discovery against tuberculosis from natural products.
Currently, one third of the world's population is latently infected with Mycobacterium tuberculosis (MTB), while 8.9-9.9 million new and relapse cases of tuberculosis (TB) are reported yearly. The renewed research interests in natural products in the hope of discovering new and novel antitubercular leads have been driven partly by the increased incidence of multidrug-resistant strains of MTB and the adverse effects associated with the first- and second-line antitubercular drugs. Natural products have been, and will continue to be a rich source of new drugs against many diseases. The depth and breadth of therapeutic agents that have their origins in the secondary metabolites produced by living organisms cannot be compared with any other source of therapeutic agents. Discovery of new chemical molecules against active and latent TB from natural products requires an interdisciplinary approach, which is a major challenge facing scientists in this field. In order to overcome this challenge, cutting edge techniques in mycobacteriology and innovative natural product chemistry tools need to be developed and used in tandem. The present review provides a cross-linkage to the most recent literature in both fields and their potential to impact the early phase of drug discovery against TB if seamlessly combined.


Posterior auricular artery as an alternative donor vessel for extracranial-intracranial bypass surgery.
Sometimes the superficial temporal artery (STA) is not available for an extracranial-intracranial (EC-IC) bypass procedure. An alternative vessel for an EC-IC bypass is the posterior auricular artery (PAA) if it extends to the temporoparietal area with a diameter large enough. We assessed the prevalence of an appropriate PAA as an alternative donor vessel and report three illustrative cases in which the PAA was used for EC-IC bypass surgery. A literature search was performed on the use of the PAA as a donor vessel for bypass surgery. Secondly, a prospective database of bypass surgeries was reviewed to calculate the prevalence of a PAA with a diameter of at least 1 mm in the parietotemporal area. Finally, three illustrative cases are reported that describe various indications for the revascularisation procedures with their clinical, surgical and imaging features. Two articles have previously described the use of the PAA for bypass surgery and their results are summarised. The prevalence of a PAA that would be appropriate for an EC-IC bypass in patients with intracranial vascular pathology is 5.7%. The presented cases demonstrate that the PAA can be successfully used for EC-IC bypass surgery with good flow velocities and patency. The PAA is a rarely described as an appropriate donor vessel for an EC-IC bypass. Its prevalence is 5.7% and it can successfully be used as an alternative donor vessel. The awareness among cerebrovascular surgeons about the presence of a PAA and knowledge about its anatomy may be valuable.


Cancer communication and informatics research across the cancer continuum.
Over the past decade, dramatic changes brought about by a rapid diffusion of Internet technologies, cellular telephones, mobile devices, personal digital assistants, electronic health records, and data visualization have helped to create a revolution in health communication. To understand the implications of this communication revolution for cancer care, the National Cancer Institute launched an ambitious set of research priorities under its "extraordinary opportunities" program. We present an overview of some of the relevant behavioral research being conducted within the perspective of this extraordinary opportunity in cancer communication research. We begin by tracing the implications of this research for behavioral scientists across the continuum of cancer care from primary prevention (e.g., tobacco control, diet, exercise, sun protection, and immunization against human papilloma virus), to secondary prevention (e.g., screening for polyps, lesions, and early stage neoplasms), to diagnosis and treatment, posttreatment survivorship, and end of life. Along each point of the continuum, we describe a natural evolution of knowledge from studies on the traditional role of media to research on the changing role of new media and informatics, and we carefully highlight the role that psychological research has played in improving communication- and health-related outcomes along the way. We conclude with an appeal to psychologists of many different backgrounds to join with biomedical researchers, engineers, clinical practitioners, and others to accelerate progress against cancer.


Commercial tattooing as a potentially important source of hepatitis C infection. Clinical epidemiology of 626 consecutive patients unaware of their hepatitis C serologic status.
Tattooing in commercial tattoo parlors is known to transmit blood-borne viral infections, including hepatitis C virus (HCV), in other countries, but its contribution to the high population prevalence of HCV infection in the United States has been incompletely evaluated. Risk factors for blood-borne infection were assessed by physician's interview of 626 consecutive patients undergoing medical evaluation for spinal problems in 1991 and 1992 while unaware of their HCV status. Later all were screened for HCV infection with enzyme-linked immunosorbent assay (EIA-1 and EIA-2), and positives were confirmed with second-generation recombinant immunoblot assay (RIBA). Forty-three patients were seropositive for HCV (sample prevalence 6.9%, population-standardized prevalence 2.8%). Logistic regression analysis identified 4 independent risk factors for HCV infection: injection-drug use (adjusted prevalence odds ratio [OR] = 23.0; 95% confidence intervals [CI] = 7.5-70.6), ancillary hospital jobs held by men (OR = 9.6; 95% CI = 3.8-24.3), tattoos from commercial tattoo parlors (OR = 6.5; 95% CI = 2.9-14.8), and drinking > or = 3 6-packs of beer per month (OR = 4.0; 95% CI = 1.8-8.7). If causal, these 4 risk factors account for 91% of HCV infections, with tattooing explaining 41%, heavy beer drinking 23%, injection-drug use 17%, and ancillary health care jobs for men 8%. Transfusions, promiscuous sexual activity, bone grafts, acupuncture, perinatal or intimate transmission in families, and other modes were not independently associated with serologic evidence of HCV infection. Unlikely to be explained by confounding or incomplete disclosure of other risk factors, tattooing in commercial tattoo parlors may have been responsible for more HCV infections than injection-drug use.


Pre-pregnancy adherences to empirically derived dietary patterns and gestational diabetes risk in a Mediterranean cohort: the Seguimiento Universidad de Navarra (SUN) project.
The aim of this study was to investigate the association between the adherence to empirically derived dietary patterns and gestational diabetes mellitus (GDM) risk and of healthy lifestyles with the prevention of GDM defining an overall healthy score. The Seguimiento Universidad de Navarra project is a Mediterranean cohort of university graduates started in 1999. We included 3455 pregnant women. During a mean follow-up of 10·3 (sd 3·3) years, we identified 173 incident GDM cases. Two major dietary patterns were identified using principal component analysis: the Western dietary pattern (WDP) (characterised by a high consumption of meat-based products and processed foods) and the Mediterranean dietary pattern (MDP) (characterised by a high consumption of vegetables, fruits, fish and non-processed foods). A low-risk score for GDM was defined taking into account important risk factors (age, BMI and unhealthy dietary pattern) for GDM. Positive association was found in the multivariable model between the highest quartile of adherence to WDP and GDM incidence compared with the lowest quartile (OR 1·56; 95 % CI 1·00, 2·43). No association was found between adherence to the MDP and GDM incidence (OR 1·08; 95 % CI 0·68, 1·70 for the highest quartile compared with the lowest). Women who adhered to all three low-risk factors had a 76 % lower risk of GDM (OR 0·24; 95 % CI 0·10, 0·55) compared with women who did not adhere to any factor before pregnancy. In conclusion, our results reinforce the importance of dietary recommendations and other two factors (low BMI and young age at pregnancy) in pre-gravid women.


Integration of acupuncture for outpatients and inpatients in a general hospital in Brazil.
Acupuncture and related techniques have increasingly been offered in conventional medical settings in Western societies. In Hospital Israelita Albert Einstein, Brazil, acupuncture has been integrated into the care pathways since October 2005. Since then, medical acupuncture has been offered for both inpatients and outpatients. Acupuncture has become an integrated therapeutic modality both for outpatients in the clinic setting and for inpatients in the wards. It has been observed that acupuncture performed in a hospital differs in specific characteristics when compared with acupuncture performed in an outpatient setting. The main differences found between inpatients and outpatients treatment are summarised and attitudes and cautionary measures to be taken into account during application of acupuncture in inpatients are suggested. Future plans for the service include offering acupuncture in the emergency ward and surgical centre. The description of this experience could encourage other hospitals to develop an acupuncture service.


Inhibition of human cytosolic phospholipase A2 by human annexin V.
The ability of annexins, particularly annexin 1 (lipocortin 1), to inhibit phospholipase A2 (PLA2) is well known and a substrate depletion mechanism is now widely accepted as the explanation for most inhibitory studies. However, there are only a very limited number of reported studies involving annexins and the high-molecular-mass cytosolic PLA2 (cPLA2). In this study we have examined the effect of human recombinant annexin V, a potentially abundant cytosolic protein, on the ability of human recombinant cPLA2 to hydrolyse a variety of phospholipid substrates. The results show clearly that, under the conditions of our study, annexin V can inhibit cPLA2 activity by a mechanism of substrate depletion and that this inhibition is dependent on the nature of the phospholipids and the concentration of Ca2+ ions in the assay. The hydrolysis of 1-stearoyl 2-arachidonyl phosphatidylcholine by cPLA2 was not significantly affected by annexin V over a range of Ca2+ concentrations (1 microM-2.5 mM), a result that presumably reflects the zwitterionic nature of the phospholipid and the known inability of annexins to bind to such interfaces. In contrast, the hydrolysis of dioleoyl phosphatidylglycerol, which is an effective anionic phospholipid substrate for this enzyme, and more significantly that of 1-stearoyl 2-arachidonyl phosphatidic acid, were readily inhibited by annexin V, although these effects were Ca2+-dependent. The Ca2+ concentrations required for inhibition in the assay system in vitro are greater than those associated with Ca2+-stimulated events within the cell, suggesting that a role for annexin V in regulating cPLA2 activity might not involve a substrate depletion mechanism in vivo unless factors in addition to Ca2+ and phospholipids contribute to the binding of annexin V to cell membranes.


Changes in oxygenation in mechanically ventilated critically ill patients following hyperbaric treatment.
Some ventilated intensive care unit (ICU) patients may experience reduced oxygenation following hyperbaric oxygen treatment (HBOT). In a prospective, single-centre, observational study, we documented changes in oxygenation and the need for associated changes in ventilator settings in 25 consecutive, mechanically ventilated ICU patients immediately post-treatment and 1, 2, 3 and 6 hours following 61 HBOT sessions. The primary outcome measure of oxygenation was the ratio of arterial partial pressure of oxygen (P(a)O2) against the level of inspired oxygen (F(i)O2), P(a)O2/F(i)O2. Following HBOT, the P(a)O2/F(i)O2 ratio decreased by 27% on return to ICU (P < 0.001, 95% confidence intervals (CI) 20.6 to 34.2); 22% at 1 hour post-HBOT (P < 0.001, 95% CI 15.1 to 28.6); and 8% at 2 hours post (P = 0.03, 95% CI 0.8 to 14.4). The ratio showed no significant differences from pre-HBOT at 3 and 6 hours post-HBOT. P(a)O2/F(i)O2 ratio changes necessitated adjustments to ventilation parameters upon return to ICU following 30 of 61 HBOT sessions in 17 out of the 25 patients. The most common ventilation parameter altered was F(i)O2 (n = 20), increased by a mean of +0.17 (95% CI 0.11 to 0.23) above baseline for two hours following HBOT. Following HBOT, oxygenation is reduced in a majority of mechanically ventilated ICU patients and requires temporary alterations to mechanical ventilation settings. Further study to identify predictive characteristics and to determine causation for those at risk of needing ventilation alterations is required.


Diverse roles of TssA-like proteins in the assembly of bacterial type VI secretion systems.
Protein translocation by the bacterial type VI secretion system (T6SS) is driven by a rapid contraction of a sheath assembled around a tube with associated effectors. Here, we show that TssA-like or TagA-like proteins with a conserved N-terminal domain and varying C-terminal domains can be grouped into at least three distinct classes based on their role in sheath assembly. The proteins of the first class increase speed and frequency of sheath assembly and form a stable dodecamer at the distal end of a polymerizing sheath. The proteins of the second class localize to the cell membrane and block sheath polymerization upon extension across the cell. This prevents excessive sheath polymerization and bending, which may result in sheath destabilization and detachment from its membrane anchor and thus result in failed secretion. The third class of these proteins localizes to the baseplate and is required for initiation of sheath assembly. Our work shows that while various proteins share a conserved N-terminal domain, their roles in T6SS biogenesis are fundamentally different.


Polymorphisms in glutathione S-transferase omega-1 gene and increased risk of sporadic Alzheimer disease.
Previous studies examining the association between the glutathione S-transferase omega-1 (GSTO1) single-nucleotide polymorphisms (SNPs) and Alzheimer disease (AD) have yielded conflicting results. Furthermore, an effect of GSTO1 rs4925 on the age-at-onset (AAO) of AD was found in different studies on sporadic and familial AD cases, but with contrasting findings. A total sample of 103 AD patients, and 157 age- and sex-matched unrelated caregivers from Apulia, southern Italy, were genotyped for the apolipoprotein E (APOE) polymorphism and the GSTO1 rs4925 and rs1804834 SNPs. Furthermore, we performed a haplotype analysis on these two SNPs on the GSTO1 locus and evaluated the possibility of interaction with APOE. Significant differences were observed in rs4925 genotype distribution between AD patients and age- and sex-matched healthy controls. Both the C/A (odds ratio [OR] = 3.116; 95% confidence interval [CI], 1.749-5.550) and the A/A (OR = 10.802; 95% CI, 3.605-32.128) genotypes resulted in an association with AD. A higher frequency of the allele A was observed in AD patients than in age- and sex-matched controls (OR = 3.789; 95% CI, 2.442-5.878). No significant differences were observed in the rs1804834 genotype or allele frequencies between AD patients and controls. No significant influence of the GSTO1 genotypes on the AAO was observed. No significant interaction was found among the GSTO1 SNPs and APOE. In both AD and controls, no important linkage disequilibrium (LD) was observed among the markers investigated. Whereas the C-A haplotype appeared to be protective against AD (OR = 0.303; 95% CI, 0.204-0.451), the A-A haplotype appeared to be at increased risk for AD (OR = 4.014,; 95% CI, 2.528-6.382). Our findings supported a role of the GSTO1 rs4925 SNP in the risk of sporadic AD in southern Italy, suggesting that this and other variants of the GSTO1 gene could be implicated in AD pathogenesis.


How do latent print examiners perceive proficiency testing? An analysis of examiner perceptions, performance, and print quality.
Proficiency testing has the potential to serve several important purposes for crime laboratories and forensic science disciplines. Scholars and other stakeholders, however, have criticized standard proficiency testing procedures since their implementation in laboratories across the United States. Specifically, many experts label current proficiency tests as non-representative of actual casework, at least in part because they are not sufficiently challenging (e.g., [1-4]. In the current study, we surveyed latent print examiners (n = 322) after they completed a Collaborative Testing Services proficiency test about their perceptions of test items. We also evaluated respondents' test performance and used a quality metric algorithm (LQMetrics) to obtain objective indicators of print quality on the test. Results were generally consistent with experts' concerns about proficiency testing. The low observed error rate, examiner perceptions of relative ease, and high objective print quality metrics together suggest that latent print proficiency testing is not especially challenging. Further, examiners indicated that the test items that most closely resembled real-world casework were also the most difficult and contained prints of the lowest quality. Study findings suggest that including prints of lower quality may increase both the difficulty and representativeness of proficiency testing in latent print examination.


An alternative database for the computerised surgical logbook.
A computerised surgical logbook has many advantages, but some may find the prospect of database design daunting. In this paper the author hopes, by description of his own logbook, to encourage the computer-wary. Jargon is explained, and the fundamental steps necessary to create a computerised logbook are outlined. Consideration is given to coding of operative procedure, and to the future of computerised logbooks in training.


Immunization against diseases caused by Theileria parva: a review.
Theileria parva is the causative agent of three epidemiologically different diseases, East Coast fever (ECF), Corridor disease and January disease, caused by 3 types of T. parva, T. p. parva, T. p. lawrencei and T. p. bovis, respectively. The history of immunization against these diseases has been marked by salient discoveries such as the immune status in recovered animals, the activity of tetracyclines during the incubation period, the possibility for cryopreserving supernatant of prefed ticks and the development of useful serological tests. The possibility of simultaneous administration of stabilate and long-acting tetracycline have greatly contributed to making the infection and treatment method operational. The importance of antigenic diversity in T. parva has been reflected in the difficulties related to the selection of the immunizing stock or combinations of stocks: a 'cocktail' of East African isolates may give broad protection against field challenge by ECF (T. parva parva), but Corridor disease is more problematic. On the other hand, certain single isolates may give equally good protection against ECF field challenge. Studies on the immunology of T. parva infection and the application of molecular tools have led to the discovery that sera of recovered animals neutralize sporozoites of various isolates, and to the p67 molecular vaccine; yet so far the only available method of immunizing against T. parva infections is the infection and treatment method or, in the case of T. parva bovis, the use of sublethal stabilate doses. Infection and treatment is applied on a fairly large scale in Zambia, and on a more limited scale in a few other countries. Immunity by this rather crude method is long-lasting and solid, but cross-immunity problems against some field strains remain. Furthermore, as immunized animals remain carriers, immunization may contribute to attaining and improving endemic stability in endemic areas in indigenous breeds with an adequate level of genetic tolerance to ECF. On the other hand, carrier animals may constitute a risk for spreading the disease into ECF-free areas where the vector is present. Other disadvantages of the method are that immunization of cattle during the incubation of naturally contracted East Coast fever will not prevent the disease and jeopardize its reputation. Furthermore, stabilates have to be cryopreserved, often a technical drawback, and contamination with undesirable pathogens may occur in tick-derived material. Therefore the need remains for the development of effective molecular vaccines and it must be remembered that immunization must be cost-effective and sustainable and it is only one aspect of integrated control of theileriosis and other tick-borne diseases. There is no universally valid strategy. Several factors have to be considered: value and susceptibility of cattle to theileriosis and to other tick-borne and tick-associated diseases, infestation by various ticks present in the area, the type of theileriosis (ECF, Corridor disease or January disease) and the epidemiological situation where immunization is taking place. The optimal age for immunization of the calves in endemic areas needs to be determined: when calf mortality by naturally occurring theileriosis is a problem, the sooner calves are immunized the better, but a proportion will have contracted natural infection before they can be reached, and immunization of very young calves might not be fully effective.


Impact of the ultrarapid CYP2C19*17 allele on serum concentration of escitalopram in psychiatric patients.
Recently, a novel allelic variant of cytochrome P450 2C19 encoding ultrarapid enzyme activity was described (denoted CYP2C19*17). The objective of this study was to evaluate the impact of CYP2C19*17 on serum concentration of escitalopram in psychiatric patients. One hundred and sixty-six patients treated with escitalopram were divided into the following subgroups according to CYP2C19 genotype: CYP2C19*17/*17 (n=7), CYP2C19*1/*17 (n=43), CYP2C19*1/*1 (n=60), CYP2C19*17/def (n=16), CYP2C19*1/def (n=34), and CYP2C19def/def (n=6) (def=defective allele, i.e., CYP2C19*2 or *3). Dose-adjusted serum concentrations of escitalopram were compared using the CYP2C19*1/*1 subgroup as reference. Geometric mean of the escitalopram serum concentration was 42% lower in patients homozygous for CYP2C19*17 (P<0.01) and 5.7-fold higher in subjects homozygous for defective CYP2C19 alleles (P<0.001). Of the heterozygous subgroups, only CYP2C19*1/def was significantly different from CYP2C19*1/*1 (P<0.001). In conclusion, a homozygous CYP2C19*17 genotype is associated with lower serum concentration of escitalopram, which might imply increased risk of therapeutic failure.


Mouse blood monocytes: standardizing their identification and analysis using CD115.
Monocytes have been used to assess immune dysfunction and disease. While mouse models are a useful longitudinal analog, few researchers have assessed changes in mouse monocytes. The purpose of this study was to provide recommendations for the sample processing and flow cytometric analysis of mouse blood monocytes. Blood was drawn in a non-lethal manner from CD-1 male mice to be used in three experiments. Experiment 1 compared commonly used mouse monocyte markers. Experiment 2 compared the stability of CD115 expression after immediate (0h) and delayed (2 and 4h) processing following blood collection under various experimental conditions (laser strength, anticoagulant, and storage temp.). Experiment 3 compared the consistency of CD115(+) monocyte and subset concentrations using decreasing (40, 20, 10 and 5μL) volumes of blood. In experiment 1, >95% of CD115(+) events co-expressed CD11b; >85% co-expressed CD14. 70% of CD14(+) and 50% of CD11b(+) events co-expressed CD115. In experiment 2, CD115 expression decreased by 33% between 0 and 4h when stored at room temperature. Blood treated with EDTA and refrigerated maintained CD115 stability. In experiment 3, calculated concentrations for total monocyte events varied by <10% when 40, 20 and 10μL of blood were stained. While CD115 staining provides the most distinct monocyte population, it is important to treat blood with EDTA and refrigerate if sample processing will be delayed over 2h. Collectively, the findings of the present study outline important considerations that must be addressed when examining mouse monocytes in small, non-lethal blood samples.


The breakage-fusion-bridge (BFB) cycle as a mechanism for generating genetic heterogeneity in osteosarcoma.
Osteosarcoma (OS) is characterized by chromosomal instability and high copy number gene amplification. The breakage-fusion-bridge (BFB) cycle is a well-established mechanism of genome instability in tumors and in vitro models used to study the origins of complex chromosomal rearrangements and cancer genome amplification. To determine whether the BFB cycle could be increasing the de novo rate of formation of cytogenetic aberrations in OS, the frequency of anaphase bridge configurations and dicentric chromosomes in four OS cell lines was quantified. An increased level of anaphase bridges and dicentrics was observed in all the OS cell lines. There was also a strong association between the frequencies of anaphase bridges, dicentrics, centrosomal anomalies, and multipolar mitotic figures in all the OS cell lines, indicating a possible link in the mechanisms that led to the structural and numerical instabilities observed in OS. In summary, this study has provided strong support for the role of the BFB cycle in generating the extensive structural chromosome aberrations, as well as cell-to-cell cytogenetic variation observed in OS, thus conferring the genetic diversity for OS tumor progression.


Importance of cloud motion on cigarette smoke deposition in lung airways.
Deposition patterns of mainstream cigarette smoke were studied in casts of human extrathoracic and lung airways. The laboratory tests were designed to simulate smoking (i.e., the behavior of undiluted cigarette smoke in smokers' lungs), not secondary exposures to non-smokers. The experimental data revealed concentrated deposits at well-defined sites, particularly at bifurcations (most notably at inclusive carinal ridges) and certain segments of tubular airways. The measurements suggest the occurrence of cloud motion wherein particles are not deposited by their individual characteristics but behave as an entity. The observed behavior is consistent with the theory of Martonen (1992), where it was predicted that cigarette smoke could behave aerodynamically as a large cloud (e.g., 20 microns diameter) rather than as submicrometer constituent particles. The effects of cloud motion on deposition are pronounced. For example, an aerosol with a mass median aerodynamic diameter (MMAD) of 0.443 micron and geometric standard deviation (GSD) of 1.44 (i.e., published cigarette smoke values) will have the following deposition fractions: lung (TB + P) = 0.14, tracheobronchial (TB) = 0.03, and pulmonary (P) = 0.11. When cloud motion is simulated, total deposition increases to 0.99 and is concentrated in the TB compartment, especially the upper bronchi; pulmonary deposition is negligible. Cloud motion produces heterogeneous deposition resulting in increased exposures of underlying airway cells to toxic and carcinogenic substances. The deposition sites correlated with incidence of cancers in vivo. At present, cloud motion concentration effects per se are not addressed in federal regulatory standards. The experimental and theoretical data suggest that concentrations of particulate matter may be an important factor to be integrated into U.S. Environmental Protection Agency (EPA) risk assessment protocols.


Gonadotropin-releasing hormone receptors.
GnRH and its analogs are used extensively for the treatment of hormone-dependent diseases and assisted reproductive techniques. They also have potential as novel contraceptives in men and women. A thorough delineation of the molecular mechanisms involved in ligand binding, receptor activation, and intracellular signal transduction is kernel to understanding disease processes and the development of specific interventions. Twenty-three structural variants of GnRH have been identified in protochordates and vertebrates. In many vertebrates, three GnRHs and three cognate receptors have been identified with distinct distributions and functions. In man, the hypothalamic GnRH regulates gonadotropin secretion through the pituitary GnRH type I receptor via activation of G(q). In-depth studies have identified amino acid residues in both the ligand and receptor involved in binding, receptor activation, and translation into intracellular signal transduction. Although the predominant coupling of the type I GnRH receptor in the gonadotrope is through productive G(q) stimulation, signal transduction can occur via other G proteins and potentially by G protein-independent means. The eventual selection of intracellular signaling may be specifically directed by variations in ligand structure. A second form of GnRH, GnRH II, conserved in all higher vertebrates, including man, is present in extrahypothalamic brain and many reproductive tissues. Its cognate receptor has been cloned from various vertebrate species, including New and Old World primates. The human gene homolog of this receptor, however, has a frame-shift and stop codon, and it appears that GnRH II signaling occurs through the type I GnRH receptor. There has been considerable plasticity in the use of different GnRHs, receptors, and signaling pathways for diverse functions. Delineation of the structural elements in GnRH and the receptor, which facilitate differential signaling, will contribute to the development of novel interventive GnRH analogs.


Paroxysmal nocturnal hemoglobinuria and pregnancy before the eculizumab era: the French experience.
Pregnancy in women with paroxysmal nocturnal hemoglobinuria is rare, with few reports on maternal and fetal mortality rates. A specific questionnaire designed to solicit data on pregnancies in women with paroxysmal nocturnal hemoglobinuria was sent to all members of the French Society of Hematology in January 2008. We identified 27 pregnancies in 22 women at 10 French Society of Hematology centers between 1978 and 2008. The median age was 21.5 years at diagnosis of paroxysmal nocturnal hemoglobinuria and 27 years at pregnancy. None of these women had received eculizumab during their pregnancy. Maternal complications, consisting mostly of cytopenias requiring transfusions, occurred in 95% of cases. Two cases of severe aplastic anemia (de novo in one case and relapse in the other) were recorded. No thrombotic events occurred during pregnancy, whereas 4 postpartum thromboses (16%) were recorded, 2 of which were fatal (maternal mortality rate 8%). Most patients received antithrombotic prophylaxis during pregnancy and postpartum (n=16; 64%). Delivery was preterm in 29% of cases, and birth weight was less than 3 kg in 53% of cases. Fetal mortality rate was 4%. Pregnancy during paroxysmal nocturnal hemoglobinuria is associated with increased maternal and fetal mortality rates (8% and 4%, respectively, in this series). Maternal mortality is related to postpartum thromboses. Prophylactic anticoagulation is recommended during pregnancy and for six weeks postpartum.


Wear of congruent meniscal bearings in unicompartmental knee arthroplasty: a retrieval study of 16 specimens.
Many designs of unicompartmental knee replacement show early and mid-term failure due to polyethylene wear. We studied the wear rate of congruent polyethylene meniscal bearings retrieved from failed Oxford unicompartmental knee replacements. We examined 16 bearings, 0.8 to 12.8 years after implantation, measuring their thickness and comparing it with that of 14 unused bearings. The mean rate of penetration, which included the effects of wear at both upper and lower surfaces, was 0.036 mm per year (maximum 0.08). Bearings as thin as 3.5 mm wore no faster than thicker models, but ten with evidence of impingement had greater wear. The six bearings with no impingement showed a mean rate of penetration of 0.01 mm per year. In unicompartmental knee replacement, careful implantation of fully congruous meniscal bearings can avoid failure due to polyethylene wear.


Rice mitogen-activated protein kinase interactome analysis using the yeast two-hybrid system.
Mitogen-activated protein kinase (MAPK) cascades support the flow of extracellular signals to intracellular target molecules and ultimately drive a diverse array of physiological functions in cells, tissues, and organisms by interacting with other proteins. Yet, our knowledge of the global physical MAPK interactome in plants remains largely fragmented. Here, we utilized the yeast two-hybrid system and coimmunoprecipitation, pull-down, bimolecular fluorescence complementation, subcellular localization, and kinase assay experiments in the model crop rice (Oryza sativa) to systematically map what is to our knowledge the first plant MAPK-interacting proteins. We identified 80 nonredundant interacting protein pairs (74 nonredundant interactors) for rice MAPKs and elucidated the novel proteome-wide network of MAPK interactors. The established interactome contains four membrane-associated proteins, seven MAP2Ks (for MAPK kinase), four MAPKs, and 59 putative substrates, including 18 transcription factors. Several interactors were also validated by experimental approaches (in vivo and in vitro) and literature survey. Our results highlight the importance of OsMPK1, an ortholog of tobacco (Nicotiana benthamiana) salicyclic acid-induced protein kinase and Arabidopsis (Arabidopsis thaliana) AtMPK6, among the rice MAPKs, as it alone interacts with 41 unique proteins (51.2% of the mapped MAPK interaction network). Additionally, Gene Ontology classification of interacting proteins into 34 functional categories suggested MAPK participation in diverse physiological functions. Together, the results obtained essentially enhance our knowledge of the MAPK-interacting protein network and provide a valuable research resource for developing a nearly complete map of the rice MAPK interactome.


[Lung and mediastinum puncture under ultrasound guidance].
Based on their own experience with punctures under the guidance of ultrasound study of the bulky formations of the lung and mediastinum in 254 patients, the authors show the potentialities of this technique. The paper describes the technique and its efficiency, shows difficulties and possible complications due to its use.


Clostridium chromiireducens sp. nov., isolated from Cr(VI)-contaminated soil.
A Cr(VI)-resistant, Gram-positive, spore-forming, obligate anaerobe, designated GCAF-1(T), was isolated from chromium-contaminated soil by its ability to reduce Cr(VI) in low concentrations. Mixed acid fermentation during growth on glucose resulted in accumulation of acetate, butyrate, formate and lactate. Morphological studies indicated the presence of peritrichous flagella, pili and an S-layer. The major cellular fatty acids (>5 %) were C(16 : 0), C(14 : 0), summed feature 3 (comprising iso-C(15 : 0) 2-OH and/or C(16 : 1)ω7c), C(18 : 1)ω7c, C(16 : 1)ω9c, summed feature 4 (comprising iso-C(17 : 1) I and/or anteiso-C(17 : 1) B) and C(18 : 1)ω9c. The DNA G+C content of strain GCAF-1(T) was 30.7 mol%. Phylogenetic interference indicated that strain GCAF-1(T) clustered with group I of the genus Clostridium. Of strains within this cluster, strain GCAF-1(T) shared the highest 16S rRNA gene sequence similarities (98.1-98.9 %) with Clostridium beijerinckii DSM 791(T), C. saccharobutylicum NCP 262(T), C. saccharoperbutylacetonicum N1-4(T), C. puniceum DSM 2619(T) and C. roseum DSM 51(T). However, strain GCAF-1(T) could be clearly distinguished from its closest phylogenetic neighbours by low levels of DNA-DNA relatedness (<50 %) and some phenotypic features. Based on the evidence presented here, strain GCAF-1(T) ( = DSM 23318(T) = KCTC 5935(T)) represents a novel species of the genus Clostridium, for which the name Clostridium chromiireducens sp. nov. is proposed.


Avian pancreatic polypeptide phase shifts hamster circadian rhythms when microinjected into the suprachiasmatic region.
The suprachiasmatic nucleus has been identified tentatively as a circadian pacemaker. To examine the functional role of peptides found within suprachiasmatic neurons, avian pancreatic polypeptide and vasopressin were microinjected into the suprachiasmatic region. Avian pancreatic polypeptide, but not vasopressin, shifted the phase of the wheelrunning rhythm as a function of the time of its injection within the circadian cycle. Avian pancreatic polypeptide or a similar peptide may be one component of the neurochemical processes underlying entrainment to the light-dark cycle.


[Adult asthma exacerbations in questions].
In this article a French working party critically review the international literature to revise the definition, pathophysiology, treatment and cost of exacerbations of adult asthma. The various guidelines do not always provide a consistent definition of exacerbations of asthma. An exacerbation can be defined as deterioration of clinical and/or functional parameters lasting more than 24 hours, without return to baseline, requiring a change of treatment. No single clinical or functional criterion can be used as an early marker of an exacerbation. Innate and acquired immune mechanisms, modified by contact with infectious, irritant or allergenic agents, participate in the pathogenesis of exacerbations, which are accompanied by bronchial inflammation. In 2010, mortality is related to progression of exacerbations, often occurring before the patient seeks medical attention. The objective of treatment is to control asthma and prevent exacerbations. However, many factors can trigger exacerbations and often cannot be controlled. The efficacy of inhaled corticosteroids has been demonstrated on reduction of the number of exacerbations and the number of asthma-related deaths. This treatment is cost-effective, especially in terms of reduction of exacerbations.


Comparison of the prebiotic properties of native chicory and synthetic inulins using swine fecal cultures.
Inulin-type fructans are known to exert different effects on the fermentation profile depending on the average and range of the degree of polymerization (DP). Here, swine fecal cultures were used to investigate the prebiotic properties of native chicory inulin (NIN), extracted from the chicory root, and synthetic inulin (SIN), which has a narrower DP distribution than NIN. Both NIN and SIN showed prebiotic effects, but NIN exhibited a significant decrease in pH and increase in the production of propionate and butyrate compared to SIN. There were also differences in the production of succinate and lactate, the precursors of propionate and butyrate, and the relative abundance of associated genes. Furthermore, NIN induced the growth of certain species of Bifidobacterium and Lactobacillus more strongly than SIN. These results suggest that NIN and SIN exhibit different prebiotic properties due to differences in DP, and that NIN might be more beneficial to host health.


Midline Compared With Peripheral Intravenous Catheters for Therapy of 4 Days or Longer in Pediatric Patients: A Randomized Clinical Trial.
Peripheral intravenous catheters (PIVCs) frequently fail during treatment causing therapy interruption, pain, recatheterization, and additional health care costs. Midline catheters (MCs) may improve functional dwell time and reduce failure compared with traditional PIVCs. To compare device failure of MCs with PIVCs. This was a pragmatic, randomized clinical superiority trial with an embedded internal pilot study conducted from July 2020 to May 2022. The study took place in a quaternary pediatric hospital in Brisbane, Queensland, Australia. Inclusion criteria were patients aged 1 to 18 years requiring peripherally compatible intravenous therapy for 4 days or longer. Patients were randomly assigned 1:1 to receive a PIVC or MC, stratified by age (≤5 years, >5 years). One catheter was studied per patient. The primary outcome was all-cause device failure, defined as premature cessation of device function. Secondary outcomes included number of insertion attempts, insertion failure, pain (on insertion), procedural time, patient/parent satisfaction (with insertion), device dwell time, device complications during dwell time, additional vascular access devices required to complete treatment, clinician satisfaction (at removal), and health care costs. Of the 128 patients randomly assigned to study groups, 127 patients (median [IQR] age, 7 [2-13] years; 71 male [56%]) had a device inserted, with 65 (51.2%) in the PIVC group and 62 (48.8%) in the MC group. All patients were included in the intention-to-treat analysis. Device failure was lower in patients in the MC group (10 [16.1%]) compared with those in the PIVC group (30 [46.2%]; odds ratio [OR], 0.22; 95% CI, 0.10-0.52; P <.001). MCs were associated with fewer insertion attempts (mean difference [MD], -0.3; 95% CI, -0.5 to 0; P = .04), increased dwell time (MD, 66.9 hours; 95% CI, 36.2-97.5 hours; P <.001), and fewer patients required additional vascular access devices to complete treatment in the MC group (4 [6.5%]) and PIVC group (19 [29.2%]; OR, 0.17; 95% CI, 0.05-0.52; P = .002). Compared with PIVCs, use of MCs was associated with greater patient (9.0 vs 7.1 of 10; P = .002) and parent (9.1 vs 8.2 of 10; P = .02) satisfaction and lower health care costs (AUS -$151.67 [US -$101.13] per person; 95% credible interval, AUS -$171.45 to -$131.90 [US -$114.20 to -$87.95]). Findings suggest that MC insertion for patients requiring peripherally compatible intravenous therapy for 4 days or longer should be prioritized to reduce the resource intensive, expensive, and burdensome sequelae of device failure. Australia New Zealand Clinical Trials Registry, ACTRN12620000724976.


The rise of oncology biosimilars: from process to promise.
Biosimilars are biologic products that are highly similar to, and have no clinically meaningful differences from, the approved originator molecule. They are poised to play an increasingly central role in cancer treatment, helping to improve access by driving down costs. Regulatory bodies have set out robust mechanisms for the approval of biosimilars, based on comprehensive and rigorous analytical and nonclinical comparisons with the originator. Product attributes (e.g., post-translational modifications) that are important to the function of the molecule must be similar between biosimilar and originator. This should be followed by a robust clinical development program, assessing pharmacokinetics, pharmacodynamics, efficacy, safety and immunogenicity. Equivalence in one indication might allow extrapolation across all the indications of the originator biologic. The recent approval of several trastuzumab biosimilars provides an example of how this process can work in practice for the benefit of patients, clinicians and payers.


Multicenter randomized controlled trial comparing the performance of 22 gauge versus 25 gauge EUS-FNA needles in solid masses.
Few randomized studies have assessed the clinical performance of 25-gauge (25G) needles compared with 22-gauge (22G) needles during endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) biopsy of intra-abdominal lesions. We aimed to compare the diagnostic yield, as well as performance characteristics of 22G versus 25G EUS biopsy needles by determining their diagnostic capabilities, the number of needle passes as well as cellularity of aspirated tissue specimen. The study is a prospective, randomized, multicenter study. Patients were referred between January 2009 and January 2010 for diagnostic EUS including EUS-guided FNA of different lesions adjacent to the upper GI tract. All patients were randomized to EUS-FNA performed with either a 22G or 25G aspiration needle. EUS-FNA was performed in 135 patients (62 patients with a 22G needle). Sensitivity and specificity of the 22G needle was 94.1% and 95.8%, respectively, and for the 25G needle 94.1% and 100%, respectively. Investigators reported better visualization and performance for the 22G needle compared to the 25G (p < 0.0001). The number of tissue slides obtained was higher for the 22G needle during the second and third needle passes (p < 0.05). We did not observe significant differences between the number and preservation status of obtained cells (p > 0.05). A significant difference was found between the two types of needles in terms of reduced visualization of the 25G needle and suboptimal performance rating. However, this did not impact on overall results since both needles were equally successful in terms of a high diagnostic yield and overall accuracy.


In situ X-ray absorption spectroscopic studies of photocatalytic oxidation of As(III) to less toxic As(V) by TiO2 nanotubes.
Arsenic in groundwater caused the black-foot disease (BFD) in many countries in the 1950-1960s. It is of great importance to develop a feasible method for removal of arsenic from contaminated groundwater in BFD endemic areas. Photocatalytic oxidation of As(III) to less toxic As(V) is, therefore, of significance for preventing any arsenic-related disease that may occur. By in situ synchrotron X-ray absorption spectroscopy, the formation of As(V) is related to the expense of As(III) disappearance during photocatalysis by TiO2 nanotubes (TNTs). Under UV/Vis light irradiation, the apparent first-order rate constant for the photocatalytic oxidation of As(III) to As(V) is 0.0148 min-1. It seems that As(III) can be oxidized with photo-excited holes while the not-recombined electrons may be scavenged with O2 in the channels of the well defined TNTs (an opening of 7 nm in diameter). In the absence of O2, on the contrary, As(III) can be reduced to As(0), to some extent. Cu(II) (CuO), as an electron acceptor, was impregnated on the TNTs surfaces in order to gain a better understanding of electron transfer during photocatalysis. It appears that As(III) can be oxidized to As(V) while Cu(II) is reduced to Cu(I) and Cu(0). The molecular-scale data are very useful in revealing the oxidation states and interconversions of arsenic during the photocatalytic reactions. This work has implications in that the toxicity of arsenic in contaminated groundwater or wastewater can be effectively decreased via solar-driven photocatalysis, which may facilitate further treatments by coagulation.


Blood lead concentrations in exposed forecourt attendants and taxi drivers in parts of South Africa.
Leaded fuel was banned in South Africa in 2006, in order to improve human health and reduce environmental pollution. Lead (Pb) has been suggested to contribute to the development of neurodegenerative disorders, and the role of respiratory exposure to Pb from petrol fumes should not be neglected in this context. In addition to Pb, petrol contains various harmful chemicals including other neurotoxic metals and hydrocarbons. Here, we investigated concentrations of Pb and other metals in blood from petrol station forecourt attendants (n = 38), taxi drivers (n = 21), and unexposed controls (n = 36). Taxi drivers and forecourt attendants were divided into three groups each, based on number of years worked. A questionnaire was designed to investigate the health status of the participants. Blood samples were collected by medical professionals and analyzed for metal concentrations by ICP-MS. A positive correlation between number of years worked and Pb blood concentrations was found. The highest Pb concentration (60.2 µg/L) was observed in a forecourt attendant who had worked 11-20 years, and the average Pb concentration in this group (24.5 µg/L) was significantly (p < 0.05) higher than in forecourt attendants who had worked 2-5 years (10.4 µg/L). Some individuals had elevated concentrations of manganese, arsenic, cadmium, chromium and cobalt, yet not significantly elevated at the group level. The blood levels of arsenic appeared to be related to smoking. Mood swings, dizziness, headaches and tiredness were reported by the workers. Blood Pb concentrations in petrol station forecourt attendants and taxi drivers exposed to leaded petrol are elevated and correlate to exposure time. A health monitoring program should be erected for all individuals working in these industries, and preventive measures should be implemented to eliminate metal exposure from petrol.


Assessment of drug-related problems and their impact on blood pressure control in patients with hypertension.
Hypertension is among the leading causes of cardiovascular disease worldwide. No previous studies have examined specifically the types of drug-related problems (DRPs) in patients with hypertension, and how these problems affect blood pressure control. So, the primary aim of this study was to investigate the prevalence and nature of DRPs in outpatients with hypertension and reveal any associations between DRPs and blood pressure control in these patients. This was a cross-sectional correlation study. A total of 200 patients who fulfilled the inclusion criteria were recruited through outpatient clinics. Patients' data were assessed to identify DRPs using a systematic evidence-based approach. Two hundred participants were recruited in this cross-sectional study. About half of the participants (48.5%) had uncontrolled hypertension. The average number of DRPs was 6.31±2.63. The main identified DRPs affecting blood pressure control included patients' lack of knowledge about their medications, lack of adherence to their medications, suboptimal efficacy of medications prescribed and the need for additional monitoring. Logistic regression analysis was performed to identify the most important predictors, and significant associations were found between poor blood pressure control and the above-identified variables except for knowledge about medication (p≤0.05). The number of DRPs among patients with hypertension is relatively high. These DRPs were associated with poor blood pressure control. The implementation of clinical pharmacy services for all patients with hypertension is strongly recommended.


AgPd nanoparticles for electrocatalytic CO2 reduction: bimetallic composition-dependent ligand and ensemble effects.
Monodisperse AgPd nanoparticles (NPs) were synthesized and studied as an efficient catalyst for electrocatalytic CO2 reduction by modulating bimetallic compositions. The mechanistic studies, based on density functional theory (DFT) calculations and environmental diffuse reflectance infrared Fourier-transform spectroscopy (DRIFTS) analysis, revealed that the incorporation of Ag in AgPd NPs can effectively weaken CO adsorption on all possible Pd surface sites (the ligand effects), and more importantly, disrupt the strongest multi-centered CO-binding sites (the ensemble effects). With properly tuned CO adsorption, which is ordinarily too strong over pure Pd, Ag15Pd85 NPs were found to be the best composition for the efficient production of CO. They deliver a unity conversion of CO2 to CO with a high mass activity of 15.2 mA mgmetal-1 at -0.8 V vs. the reversible hydrogen electrode (RHE) and high stability with minimal change in the CO faradaic efficiency (FECO) after 12 hours of operation.


Genetic continuity and change among the Indigenous peoples of California.
Before the colonial period, California harboured more language variation than all of Europe, and linguistic and archaeological analyses have led to many hypotheses to explain this diversity1. We report genome-wide data from 79 ancient individuals from California and 40 ancient individuals from Northern Mexico dating to 7,400-200 years before present (BP). Our analyses document long-term genetic continuity between people living on the Northern Channel Islands of California and the adjacent Santa Barbara mainland coast from 7,400 years BP to modern Chumash groups represented by individuals who lived around 200 years BP. The distinctive genetic lineages that characterize present-day and ancient people from Northwest Mexico increased in frequency in Southern and Central California by 5,200 years BP, providing evidence for northward migrations that are candidates for spreading Uto-Aztecan languages before the dispersal of maize agriculture from Mexico2-4. Individuals from Baja California share more alleles with the earliest individual from Central California in the dataset than with later individuals from Central California, potentially reflecting an earlier linguistic substrate, whose impact on local ancestry was diluted by later migrations from inland regions1,5. After 1,600 years BP, ancient individuals from the Channel Islands lived in communities with effective sizes similar to those in pre-agricultural Caribbean and Patagonia, and smaller than those on the California mainland and in sampled regions of Mexico.


The myeloproliferative sarcoma virus retains transforming functions after introduction of a dominant selectable marker gene.
The dominant neomycin resistance gene (neoR) was introduced into the genome of the myeloproliferative sarcoma virus (MPSV), a replication-defective retrovirus carrying the mos oncogene. The resulting selectable neoR-MPSV virus did not lose its acute transforming property, unlike the results of attempts by other groups to insert marker genes into oncogenic viruses. NeoR-MPSV DNA was used to generate infectious virus by transfection followed by rescue with Friend or Moloney murine leukaemia virus. Infection of fibroblasts with this virus resulted in morphologically transformed cells which were resistant to the neomycin analogue G418. Segregation of the two functions (transformation and G418 resistance) was not observed in more than 500 independent viral transfers to fibroblasts. Furthermore, neoR-MPSV retained the leukaemogenesis-inducing properties of the wild-type virus. Myeloproliferation and G418-resistance transfer did not segregate after passage in mice.


Role of miR-222-3p in c-Src-Mediated Regulation of Osteoclastogenesis.
MicroRNAs (miRNAs) are small non-coding RNAs that play a mostly post-transcriptional regulatory role in gene expression. Using RAW264.7 pre-osteoclast cells and genome-wide expression analysis, we identified a set of miRNAs that are involved in osteoclastogenesis. Based on in silico analysis, we specifically focused on miR-222-3p and evaluated its role in osteoclastogenesis. The results show that the inhibitor of miR-222-3p upregulated the mRNA levels of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1) and tartrate-resistant acid phosphatase (TRAP), while its mimicking agent downregulated their mRNA levels. Western blot analysis showed that its inhibitor increased the protein levels of TRAP and cathepsin K, while its mimicking agent decreased their levels. Genome-wide mRNA expression analysis in the presence and absence of receptor activator of nuclear factor κ-B ligand (RANKL) predicted c-Src as a potential regulatory target of miR-222-3p. Live cell imaging using a fluorescence resonance energy transfer (FRET) technique revealed that miR-222-3p acted as an inhibitor of c-Src activity, and a partial silencing of c-Src suppressed RANKL-induced expression of TRAP and cathepsin K, as well as the number of multi-nucleated osteoclasts and their pit formation. Collectively, the study herein demonstrates that miR-222-3p serves as an inhibitor of osteoclastogenesis and c-Src mediates its inhibition of cathepsin K and TRAP.


Biocultural diversity and crop improvement.
Biocultural diversity is the ever-evolving and irreplaceable sum total of all living organisms inhabiting the Earth. It plays a significant role in sustainable productivity and ecosystem services that benefit humanity and is closely allied with human cultural diversity. Despite its essentiality, biodiversity is seriously threatened by the insatiable and inequitable human exploitation of the Earth's resources. One of the benefits of biodiversity is its utilization in crop improvement, including cropping improvement (agronomic cultivation practices) and genetic improvement (plant breeding). Crop improvement has tended to decrease agricultural biodiversity since the origins of agriculture, but awareness of this situation can reverse this negative trend. Cropping improvement can strive to use more diverse cultivars and a broader complement of crops on farms and in landscapes. It can also focus on underutilized crops, including legumes. Genetic improvement can access a broader range of biodiversity sources and, with the assistance of modern breeding tools like genomics, can facilitate the introduction of additional characteristics that improve yield, mitigate environmental stresses, and restore, at least partially, lost crop biodiversity. The current legal framework covering biodiversity includes national intellectual property and international treaty instruments, which have tended to limit access and innovation to biodiversity. A global system of access and benefit sharing, encompassing digital sequence information, would benefit humanity but remains an elusive goal. The Kunming-Montréal Global Biodiversity Framework sets forth an ambitious set of targets and goals to be accomplished by 2030 and 2050, respectively, to protect and restore biocultural diversity, including agrobiodiversity.


Localization of the site of ventricular premature complexes by radionuclide angiographic phase imaging.
To investigate whether gated radionuclide angiographic phase imaging is useful for visually displaying the origin of ventricular premature complexes (VPCs), 82 patients were studied by gating only VPCs. The VPC "origin" by the scintigraphic method was defined as the area of earliest phase and was compared with that predicted by 12-lead electrocardiographic criteria in all patients and to invasive electrophysiologic mapping in 10. Separating the right ventricle into 3 and the left ventricle into 4 segments, the phase imaging method and the electrocardiographic criteria agreed as to ventricle of VPC origin in 69 patients (84%) and segment of origin within each ventricle in 46 (56%). When baseline ventricular wall motion was analyzed, the 2 methods agreed to the ventricle of VPC origin in 31 of 33 patients (94%) with normal wall motion, 20 of 23 (87%) with segmental wall motion abnormalities and 19 of 26 (73%) with diffuse wall motion abnormalities. Agreement between the 2 methods as to specific segmental localization of the arrhythmia focus was noted in 21 of 33 patients (64%) with normal wall motion, 11 of 23 (48%) with segmental wall motion abnormalities and 12 of 26 (46%) with diffuse hypocontractility. In the 10 patients with endocardial mapping studies, the phase imaging technique confirmed the segment of VPC origin in all 10; the electrocardiographic method was accurate in 8. Thus, gated radionuclide angiographic phase imaging methods may be of value in noninvasively defining the origin of spontaneous VPCs. The visual format allows ready interpretation of the arrhythmia origin, and there may be an advantage to this approach over electrocardiographic morphometric criteria.


Partial recovery of abnormal insula and dorsolateral prefrontal connectivity to cognitive networks in chronic low back pain after treatment.
We previously reported that effective treatment of chronic low back pain (CLBP) reversed abnormal brain structure and functional MRI (fMRI) activity during cognitive task performance, particularly in the left dorsolateral prefrontal cortex (DLPFC). Here, we used resting-state fMRI to examine how chronic pain affects connectivity of brain networks supporting cognitive functioning and the effect of treatment in 14 CLBP patients and 16 healthy, pain-free controls (scans were acquired at baseline for all subjects and at 6-months post-treatment for patients and a matched time-point for 10 controls). The main networks activated during cognitive task performance, task-positive network (TPN) and task-negative network (TNN) (aka default mode) network, were identified in subjects' task fMRI data and used to define matching networks in resting-state data. The connectivity of these cognitive resting-state networks was compared between groups, and before and after treatment. Our findings converged on the bilateral insula (INS) as the region of aberrant cognitive resting-state connectivity in patients pretreatment versus controls. These findings were complemented by an independent, data-driven approach showing altered global connectivity of the INS. Detailed investigation of the INS confirmed reduced connectivity to widespread TPN and TNN areas, which was partially restored post-treatment. Furthermore, analysis of diffusion-tensor imaging (DTI) data revealed structural changes in white matter supporting these findings. The left DLPFC also showed aberrant connectivity that was restored post-treatment. Altogether, our findings implicate the bilateral INS and left DLPFC as key nodes of disrupted cognition-related intrinsic connectivity in CLBP, and the resulting imbalance between TPN and TNN function is partially restored with treatment.


Gradient in cytoplasmic pressure in germline cells controls overlying epithelial cell morphogenesis.
It is unknown how growth in one tissue impacts morphogenesis in a neighboring tissue. To address this, we used the Drosophila ovarian follicle, in which a cluster of 15 nurse cells and a posteriorly located oocyte are surrounded by a layer of epithelial cells. It is known that as the nurse cells grow, the overlying epithelial cells flatten in a wave that begins in the anterior. Here, we demonstrate that an anterior to posterior gradient of decreasing cytoplasmic pressure is present across the nurse cells and that this gradient acts through TGFβ to control both the triggering and the progression of the wave of epithelial cell flattening. Our data indicate that intrinsic nurse cell growth is important to control proper nurse cell pressure. Finally, we reveal that nurse cell pressure and subsequent TGFβ activity in the stretched cells combine to increase follicle elongation in the anterior, which is crucial for allowing nurse cell growth and pressure control. More generally, our results reveal that during development, inner cytoplasmic pressure in individual cells has an important role in shaping their neighbors.


Childhood-Onset Leber Hereditary Optic Neuropathy-Clinical and Prognostic Insights.
To investigate the clinical and molecular genetic features of childhood-onset Leber hereditary optic neuropathy (LHON) to gain a better understanding of the factors influencing the visual outcome in this atypical form of the disease. Retrospective cohort study. We retrospectively included 2 cohorts of patients with LHON with onset of visual loss before the age of 12 years from Italy and the United Kingdom. Ophthalmologic evaluation, including best-corrected visual acuity, orthoptic evaluation, slit-lamp biomicroscopy, visual field testing, and optical coherence tomography, was considered. Patients were classified based on both the age of onset and the pattern of visual loss. A total of 68 patients were stratified based on the age of onset of visual loss: group 1 (<3 years): 14 patients (20.6%); group 2 (≥3 to <9 years): 27 patients (39.7%); and group 3 (≥9 to ≤12 years): 27 patients (39.7%). Patients in group 2 achieved a better visual outcome than those in group 3. Patients in groups 1 and 2 had better mean deviation on visual field testing than those in group 3. The mean ganglion cell layer thickness on optical coherence tomography in group 2 was higher than those in groups 1 and 3. Patients were also categorized based on the pattern of visual loss as follows: Subacute Bilateral: 54 patients (66.7%); Insidious Bilateral: 14 patients (17.3%); Unilateral: 9 patients (11.1%); and Subclinical Bilateral: 4 patients (4.9%). Children who lose vision from LHON before the age of 9 years have a better visual prognosis than those who become affected in later years, likely representing a "form frustre" of the disease.