Fetal "gallstones" are still an unsolved mystery. Case series.
Echogenic content in the fetal gallbladder is rather an incidental finding during third-trimester ultrasonography. The etiology, clinical course, and prognosis of this condition are still unclear. We highlight the demographic, clinical and ultrasound characteristics of our patients with echogenic content in the fetal gallbladder. This is a retrospective single-center study conducted at the University of Medicine and Pharmacy Cluj-Napoca, Romania, between March 2022 and April 2023. All pregnant patients who were admitted to the hospital and had echogenic content in the fetal gallbladder detected by ultrasound were identified. The clinical and ultrasonography parameters were obtained from the databases of the ultrasound units and the medical records of the patients. Four patients were found to have echogenic content in the fetal gallbladder. The mean maternal age at diagnosis was 27.5 years (range 25-31). All patients had singleton pregnancies. The mean gestational age at diagnosis was 36.5 weeks (range 35-40). In all cases, the predominant aspect of echogenic content on ultrasound was multiple hyperechogenic foci; additionally, in one case, sludge was also observed. The mean maximal length of the hyperechogenic foci was 3.375 mm (range 1.6-5.4). All fetuses were delivered at full term, either vaginally or by cesarean section. The mean weight at birth was 3082.5 g (range 2800-3450). In all four cases, the EC disappeared spontaneously at birth. Neither of the four newborns displayed digestive symptoms or complications during the follow-up. The EC is still a medical mystery since little is known about its etiology and long-term outcome. Many cases are likely to be undetected until after birth; therefore, fetal gallbladder examination should be included in the US routine. The condition appears to be benign, and usually, it improves naturally after birth. Consecutively, asymptomatic newborns with EC can be managed through a wait-and-see approach.


Modulation of microRNA expression in human T-cell development: targeting of NOTCH3 by miR-150.
Ontogenesis of T cells in the thymus is a complex process whose molecular control is poorly understood. The present study investigated microRNAs involved in human thymocyte differentiation by comparing the microRNA expression profiles of thymocytes at the double-positive, single-positive CD4(+) and single-positive CD8(+) maturation stages. Microarray analysis showed that each thymocyte population displays a distinct microRNA expression profile that reflects their developmental relationships. Moreover, analysis of small-RNA libraries generated from human unsorted and double-positive thymocytes and from mature peripheral CD4(+) and CD8(+) T lymphocytes, together with the microarray data, indicated a trend toward up-regulation of microRNA expression during T-cell maturation after the double-positive stage and revealed a group of microRNAs regulated during normal T-cell development, including miR-150, which is strongly up-regulated as maturation progresses. We showed that miR-150 targets NOTCH3, a member of the Notch receptor family that plays important roles both in T-cell differentiation and leukemogenesis. Forced expression of miR-150 reduces NOTCH3 levels in T-cell lines and has adverse effects on their proliferation and survival. Overall, these findings suggest that control of the Notch pathway through miR-150 may have an important impact on T-cell development and physiology.


MESPEUS: a database of metal coordination groups in proteins.
MESPEUS is a freely accessible database which uses carefully selected metal coordination groups found in metalloprotein structures taken from the Protein Data Bank. The database contains geometrical information of metal sites within proteins, including 40 metal types. In order to completely determine the metal coordination, the symmetry-related units of a given protein structure are taken into account and are generated using the appropriate space group symmetry operations. This permits a more complete description of the metal coordination geometry by including all coordinating atoms. The user-friendly web interface allows users to directly search for a metal site of interest using several useful options, including searching for metal elements, metal-donor distances, coordination number, donor residue group, and structural resolution. These searches can be carried out singly or in combination. The details of a metal site and the metal site(s) in the whole structure can be graphically displayed using the interactive web interface. MESPEUS is automatically updated monthly by synchronizing with the PDB database. An investigation for the Mg-ATP interaction is given to demonstrate how MESPEUS can be used to extract information about metal sites by selecting structure and coordination features. MESPEUS is available at http://mespeus.nchu.edu.tw/.


Subglottic airway becomes stable with age in the human infant larynx.
Development of the mucosal defence system and its relationship to the pathogenesis of laryngeal diseases was studied by examining the changes in the laryngeal structures at the level of subglottic larynx of 59 human infants who died within 1 year of birth. By making a transverse section at the level of the cricoid cartilage, the area and diameter of each subglottic structure were examined using an image analysis system. The results were analysed, not only by their age at death, but also based on the total development score. The areas of the whole structure, cricoid ring, subglottic mucosa and inner cavity (available area) increased in close correlation with the total development score. In contrast to the increased diameter of the inner cavity, the thickness of the mucosa was almost the same size. These results indicate the stability of the subglottic airway with advancing age in the human infant larynx.


A Classification Proposal for Adenomyosis Based on Magnetic Resonance Imaging.
There is a lack of an international consensus on adenomyosis classification that is useful for clinical practice and research. This article reviews advancements in the classification of adenomyosis, and the existing limitations. We collected a literature search from PubMed and Embase database up to March 2019. The proposed adenomyosis classification is based on magnetic resonance imaging and clinically relevant parameters. Adenomyosis is not a disease of homogeneity but is composed of multiple heterogeneous subtypes. Adenomyosis represents a spectrum of lesions, ranging from increased thickness of the junctional zone to focal or diffuse lesions involving the entire uterine wall. Potentially important parameters to be included in the classification could be affected area (internal or external adenomyosis), pattern (focal or diffuse), size or volume (myometrial involvement <1/3, <2/3, or >2/3 of uterine wall), concomitant pathologies (none, peritoneal endometriosis, ovarian endometrioma, deep infiltrating endometriosis, uterine fibroids, or others) and localization (anterior, posterior, left lateral, right lateral, or fundal). We propose a simplified classification system to monitor symptom severity against morphological types or extent of adenomyosis using the combination of previously published classifications as a starting point. More studies are needed to investigate whether this classification represents a useful tool for disease assessment in clinical practice and research.


Cervical sympathetic chain schwannoma.
Schwannomas are benign, slow-growing tumors that arise from nerves. Those originating from the sympathetic cervical chain are rare. We describe our experience with the clinical presentation, surgical management, and outcomes of patients with this pathology. Retrospective chart review of a case series in a tertiary referral center. Four cases of cervical sympathetic chain schwannomas were reviewed. Patients presented with either an asymptomatic neck mass discovered on routine physical examination (1 patient), an enlarging neck mass (2), or an acute onset of a Horner's syndrome (1). All patients underwent preoperative imaging (magnetic resonance imaging, computed tomography, or both). The location and soft-tissue characteristics of the mass, along with displacement of the carotid sheath vessels, were typical of a cervical sympathetic chain schwannoma. All patients underwent surgical excision of the mass. Postoperative Horner's syndrome was encountered in all patients. First bite syndrome was encountered in two patients. Cervical sympathetic chain schwannomas are rare tumors. Preoperative imaging characteristics facilitate the diagnosis. First bite syndrome can occur and may be debilitating postoperatively. Long-term prognosis is excellent.


CapsidMaps: protein-protein interaction pattern discovery platform for the structural analysis of virus capsids using Google Maps.
Structural analysis and visualization of protein-protein interactions is a challenging task since it is difficult to appreciate easily the extent of all contacts made by the residues forming the interfaces. In the case of viruses, structural analysis becomes even more demanding because several interfaces coexist and, in most cases, these are formed by hundreds of contacting residues that belong to multiple interacting coat proteins. CapsidMaps is an interactive analysis and visualization tool that is designed to benefit the structural virology community. Developed as an improved extension of the φ-ψ Explorer, here we describe the details of its design and implementation. We present results of analysis of a spherical virus to showcase the features and utility of the new tool. CapsidMaps also facilitates the comparison of quaternary interactions between two spherical virus particles by computing a similarity (S)-score. The tool can also be used to identify residues that are solvent exposed and in the process of locating antigenic epitope regions as well as residues forming the inside surface of the capsid that interact with the nucleic acid genome. CapsidMaps is part of the VIPERdb Science Gateway, and is freely available as a web-based and cross-browser compliant application at http://viperdb.scripps.edu.


The comparative radiopacity of Fuji IX-GP, an intermediate restorative material.
The radiopacity of intermediate restorative materials should be sufficient to enable the clinician to distinguish the material from normal and decalcified tooth structure. The purpose of this study was to determine the relative radiopacities of intermediate restorative materials, including a newly introduced high-viscosity, self-cured, condensable glass ionomer material. Radiographs were made of six intermediate restorative materials: two reinforced zinc oxide-eugenol materials (IRM and Zinroc), a conventional glass ionomer material (Ketac-fil), a synthetic resin material (Cavit), a eugenol-free zinc oxide material (Tempit), and a new, general-purpose, condensable glass ionomer material (Fuji IX-GP). Optical density was measured using a densitometer. The optical density of dentin and enamel were used for radiographic comparison. Statistical analysis revealed significant differences among materials: Cavit = IRM = Tempit > Zinroc = Fuji IX-GP > Ketac-fil = enamel > dentin (where > indicates a statistical difference at p < or = 0.05). Although not as radiopaque as some other intermediate materials tested, the radiopacity of Fuji IX-GP appears sufficient to aid diagnosis.


Studies on production and characterization of enriched urban waste composts and their influence on crops productivity.
Enriched compost produced with use of municipal solid wastes (MSW) recorded narrow C:N ratio at the end of decomposition period than municipal solid wastes decomposed without enrichers. To enhance the decomposition rate, quality of municipal solid wastes and enrichers/amendments are found very significant for production of compost. Nutrient content particularly nitrogen, phosphorus and potassium could be enhanced with addition of organic amendment/enrichers. Response of different crops for composts produced with addition of different enrichers like night soil, 25% distillery sludge and bio-fertilizers (Azospirillum sp and Bacillus sp) was conspicuous compared to the compost derived from municipal solid wastes alone with respect to increased growth and yield of crops. Among the enriched composts, night soil enriched compost significantly increased the response of potato and groundnut crops. According to farmer's opinion obtained with matrix scoring, chemical fertilizers and sheep penning are cheaper compared to pit compost or urban solid waste compost. While chemical fertilizers are considered to have adverse effects on soil more than pit compost, tank silt, sheep penning and urban solid waste. Weed infestation is associated more with urban waste than other sources. For dry land, agriculture urban waste could be useful due to good moisture holding capacity. Crop yields could be improved under low rainfall condition whenever pit compost or urban solid waste is used.


Disc herniations in the National Football League.
Retrospective analysis of a prospectively collected database. To determine the overall incidence, location, and type of disc herniations in professional football players to target treatment issues and prevention. Disc herniations represent a common and debilitating injury to the professional athlete. The NFL's (National Football League's) Sports Injury Monitoring System is a surveillance database created to monitor the league for all injuries, including injuries to the cervical, thoracic, and lumbar spine. A retrospective analysis was performed on all disc herniations to the cervical, thoracic, and lumbar spine during a 12-season period (2000–2012) using the NFL's surveillance database. The primary data points included the location of the injury, player position, activity at time of injury, and playing time lost due to injury. During the 12 seasons, 275 disc herniations occurred in the spine. In regard to location, 76% occurred in the lumbar spine and most frequently affected the L5–S1 disc. The offensive linemen were most frequently injured. As expected, blocking was the activity that caused most injuries. Lumbar disc herniations rose in prevalence and had a mean loss of playing time of more than half the season (11 games). Thoracic disc herniations led to the largest mean number of days lost overall, whereas players with cervical disc herniations missed the most practices. Disc herniations represent a significant cause of morbidity in the NFL. Although much attention is placed on spinal cord injuries, preventive measures targeting the cervical, thoracic, and lumbar spine may help to reduce the overall incidence of these debilitating injuries. N/A


Tackling the problem of HIV drug resistance.
The virally-encoded HIV-1 protease is an effective target for antiviral drugs, however, treatment for HIV infections is limited by the prevalence of drug resistant viral mutants. In this review, we describe our three-pronged approach to analyze and combat drug resistance. Understanding the molecular basis for resistance due to protease inhibitors is a key initial step in this approach. This knowledge is being employed for the design of new, improved inhibitors with high affinity for resistant mutants as well as wild type enzyme. In parallel with experimental studies of diverse mutants and inhibitory compounds, we are developing efficient algorithms to predict drug resistance phenotype from genotype data. This approach has important practical applications in the clinic where genotyping is recommended for individuals with new infections.


Prediction Model of Conversion to Dementia Risk in Subjects with Amnestic Mild Cognitive Impairment: A Longitudinal, Multi-Center Clinic-Based Study.
Patients with amnestic mild cognitive impairment (aMCI) have an increased risk of dementia. However, conversion rate varies. Therefore, predicting the dementia conversion in these patients is important. We aimed to develop a nomogram to predict dementia conversion in aMCI subjects using neuropsychological profiles. A total of 338 aMCI patients from two hospital-based cohorts were used in analysis. All patients were classified into 1) verbal, visual, or both, 2) early or late, and 3) single or multiple-domain aMCI according to the modality, severity of memory dysfunction, and multiplicity of involved cognitive domains, respectively. Patients were followed up, and conversion to dementia within 3 years was defined as the primary outcome. Our patients were divided into a training data set and a validation data set. The associations of potential covariates with outcome were tested, and nomogram was constructed by logistic regression model. We also developed another model with APOE data, which included 242 patients. In logistic regression models, both modalities compared with visual only (OR 4.44, 95% CI 1.83-10.75, p = 0.001), late compared to early (OR 2.59, 95% CI 1.17-5.72, p = 0.019), and multiple compared to single domain (OR 3.51, 95% CI 1.62-7.60, p = 0.002) aMCI were significantly associated with dementia conversion within 3 years. A nomogram incorporating these clinical variables was constructed on the training data set and validated on the validation data set. Both nomograms with and without APOE data showed good prediction performance (c-statistics ≥ 0.75). This study showed that several neuropsychological profiles of aMCI are significantly associated with imminent dementia conversion, and a nomogram incorporating these clinical subtypes is simple and useful to help to predict disease progression.


Monoclonal antibodies against Xenopus greatwall kinase.
Mitosis is known to be regulated by protein kinases, including MPF, Plk1, Aurora kinases, and so on, which become active in M-phase and phosphorylate a wide range of substrates to control multiple aspects of mitotic entry, progression, and exit. Mechanistic investigations of these kinases not only provide key insights into cell cycle regulation, but also hold great promise for cancer therapy. Recent studies, largely in Xenopus, characterized a new mitotic kinase named Greatwall (Gwl) that plays essential roles in both mitotic entry and maintenance. In this study, we generated a panel of mouse monoclonal antibodies (MAbs) specific for Xenopus Gwl and characterized these antibodies for their utility in immunoblotting, immunoprecipitation, and immunodepletion in Xenopus egg extracts. Importantly, we generated an MAb that is capable of neutralizing endogenous Gwl. The addition of this antibody into M-phase extracts results in loss of mitotic phosphorylation of Gwl, Plk1, and Cdk1 substrates. These results illustrate a new tool to study loss-of-function of Gwl, and support its essential role in mitosis. Finally, we demonstrated the usefulness of the MAb against human Gwl/MASTL.


Combined anaerobic-aerobic treatment of landfill leachates under mesophilic, submesophilic and psychrophilic conditions.
As a first step of treatment of landfill leachates (total COD--1,430-3,810 mg/l, total nitrogen 90-162 mg/l), a performance of laboratory UASB reactors has been investigated under mesophilic (30 degrees C), sub-mesophilic (20 degrees C) and psychrophilic (10 degrees C) conditions. Under hydraulic retention times (HRT) of around 7 h, when the average organic loading rates (OLR) were around 5 g COD/l/day, the total COD removal accounted for 81% (on the average) with the effluent concentrations close to anaerobic biodegradability limit (0.25 g COD/l) for mesophilic and sub-mesophilic regimes. The psychrophilic treatment conducted under the average HRT of 8 h and the average OLR of 4.22 g COD/l/day showed a total COD removal of 47% producing the effluents (0.75 g COD/l) more suitable for subsequent biological nitrogen removal. All three anaerobic regimes used for leachate treatment were quite efficient for elimination of heavy metals (Fe, Zn, Cu, Pb, Cd) by concomitant precipitation in the form of insoluble sulphides inside the sludge bed. The application of aerobic/anoxic biofilter as a sole polishing step for psychrophilic anaerobic effluents was acceptable for elimination of biodegradable COD and nitrogen approaching the current standards for direct discharge of treated wastewater.


ASS1 Overexpression: A Hallmark of Sonic Hedgehog Hepatocellular Adenomas; Recommendations for Clinical Practice.
Until recently, 10% of hepatocellular adenomas (HCAs) remained unclassified (UHCA). Among the UHCAs, the sonic hedgehog HCA (shHCA) was defined by focal deletions that fuse the promoter of Inhibin beta E chain with GLI1. Prostaglandin D2 synthase was proposed as immunomarker. In parallel, our previous work using proteomic analysis showed that most UHCAs constitute a homogeneous subtype associated with overexpression of argininosuccinate synthase (ASS1). To clarify the use of ASS1 in the HCA classification and avoid misinterpretations of the immunohistochemical staining, the aims of this work were to study (1) the link between shHCA and ASS1 overexpression and (2) the clinical relevance of ASS1 overexpression for diagnosis. Molecular, proteomic, and immunohistochemical analyses were performed in UHCA cases of the Bordeaux series. The clinico-pathological features, including ASS1 immunohistochemical labeling, were analyzed on a large international series of 67 cases. ASS1 overexpression and the shHCA subgroup were superimposed in 15 cases studied by molecular analysis, establishing ASS1 overexpression as a hallmark of shHCA. Moreover, the ASS1 immunomarker was better than prostaglandin D2 synthase and only found positive in 7 of 22 shHCAs. Of the 67 UHCA cases, 58 (85.3%) overexpressed ASS1, four cases were ASS1 negative, and in five cases ASS1 was noncontributory. Proteomic analysis performed in the case of doubtful interpretation of ASS1 overexpression, especially on biopsies, can be a support to interpret such cases. ASS1 overexpression is a specific hallmark of shHCA known to be at high risk of bleeding. Therefore, ASS1 is an additional tool for HCA classification and clinical diagnosis.


School-based screening for scoliosis: is it cost-effective?
School-based scoliosis screening was implemented in Singapore in 1981. The rationale for the programme was so that conservative treatment (bracing) can be initiated early to prevent progression of curves, avoid the complications of severe scoliosis and reduce the need for surgery. The evidence for, or against, scoliosis screening and regular follow-up remains controversial. To date, there has been no formal cost analysis of Singapore's screening programme. The aim of this paper was to examine if there are economic justifications to continue with school-based scoliosis screening. This cost-effectiveness analysis was done by comparing Singapore's existing school-based scoliosis screening and follow-up programme with the alternative of not having a programme. As the aim of the existing programme was to detect curves early, allowing bracing to be initiated and reducing the need for surgery, this analysis assumed that without the programme, students who otherwise would have received bracing and not needed surgery, would have required surgery instead. This retrospective analysis was based on School Health Service data obtained from screening 45,485 students in 1999 and 44,051 of this same cohort in 2001. Nett programme costs and health effects were computed, and a decision rule applied. The nett cost of the current mass screening programme was negative, while the nett health effects, albeit mostly intangible, positive; which made the programme an economically valuable one. Singapore's school-based scoliosis screening programme, which is implemented as part of a larger school screening and immunisation programme, is cost-effective. Cost-effectiveness may be further improved by targeting screening at high-risk groups, such as prepubertal females. More research is needed to quantify the positive health effects of scoliosis screening.


Patient-reported outcomes in high-risk HR+ /HER2- early breast cancer patients treated with endocrine therapy with or without palbociclib within the randomized PENELOPEB study.
The PENELOPEB trial investigating efficacy and safety of additional 1-year post-neoadjuvant palbociclib to standard endocrine therapy (ET) high-risk hormone receptor-positive (HR+)/human epidermal growth factor receptor 2-negative (HER2-) early breast cancer patients failed to improve invasive disease-free survival (iDFS). This analysis compared patient-reported outcomes (PROs) between treatment groups. Patients received 13 cycles of palbociclib 125 mg/day (n = 631) or placebo (n = 619) orally for 3 out of 4 weeks + ET. European Organization for Research and Treatment of Cancer Quality-of-Life Questionnaire (EORTC QLQ-C30), its breast cancer (BR23) and fatigue (FA13) modules, mood questionnaire GAD7 and European Quality of Life 5 Dimensions (EQ-5D) instruments were used for the assessment of quality of life (QoL). Repeated-measures mixed-effects models were used to evaluate differences in PRO, changes of PRO over time, and treatment-by-time interactions. 924 of 1250 patients (73.9%) completed baseline and at least one post-baseline questionnaire of all PRO instruments. General health status (GHS)/QoL based on EORTC QLQ-C30 was high in both arms (mean [SD]: palbociclib 70.1 [19.3], placebo 71.4 [18.8]) and was slightly higher in the placebo arm (LeastSquare mean difference: 0.82, p < 0.001). Higher fatigue was reported in the palbociclib arm (mean [SD]: 30.3 [23.8] vs. placebo 28.3 [22.7]; p < 0.001). No statistically significant differences were observed among FA13 physical, cognitive, and emotional fatigue subscales. Patient-reported global QoL and fatigue did not substantially change in both treatment arms. Slight differences in GHS, physical functioning, and fatigue favored the placebo arm statistically without achieving clinically meaningful thresholds.


Agricultural input shocks affect crop yields more in the high-yielding areas of the world.
The industrialization of agriculture has led to an increasing dependence on non-locally sourced agricultural inputs. Hence, shocks in the availability of agricultural inputs can be devastating to food crop production. There is also a pressure to decrease the use of synthetic fertilizers and pesticides in many areas. However, the combined impact of the agricultural input shocks on crop yields has not yet been systematically assessed globally. Here we modelled the effects of agricultural input shocks using a random forest machine learning algorithm. We show that shocks in fertilizers cause the most drastic yield losses. Under the scenario of 50% shock in all studied agricultural inputs, global maize production could decrease up to 26%, and global wheat production up to 21%, impacting particularly the high-yielding 'breadbasket' areas of the world. Our study provides insights into global food system resilience and can be useful for preparing for potential future shocks or agricultural input availability decreases at local and global scales.


Streptosporangium minutum sp. nov., isolated from garden soil exposed to microwave radiation.
The actinobacterium, strain M26T, was isolated from garden soil that was pre-treated with microwave radiation. The soil sample was collected in Roodepoort, Gauteng Province, South Africa as part of an antibiotic-screening programme. The isolate produced branched vegetative mycelium with sporangiophores bearing small sporangia ranging from 3 to 6 μm in diameter. Rapid genus identification revealed that the isolate belongs to the genus Streptosporangium. To confirm this result, the strain was subjected to polyphasic taxonomic characterisation. Chemotaxonomic characteristics were as follows: meso-DAP in the peptidoglycan, the whole-cell hydrolysate yielded madurose, predominant menaquinones were MK9 (21%), MK9(H2) (40%), MK9(H4) (31%) and MK9(H6) (3%); the polar lipid profile included an aminolipid, phosphoglycolipids, phosphatidylethanolamine, and phosphatidylmonomethylethanolamine. In addition, the fatty acid profile showed the presence of C16:0 (12.8%), C17:1ω8c (14.2%), and 10-methyl-C17:0 (15.8%). Furthermore, 16S rRNA gene sequence phylogenetic analysis showed that the strain is closely related to members of the genus Streptosporangium, which supports its classification within the family Streptosporangiaceae. Strain M26T exhibited antibiosis against a range of pathogenic bacteria, including, but not limited to Acinetobacter baumannii ATCC 19606T, Enterobacter cloacae subsp. cloacae ATCC BAA-1143, Enterococcus faecalis ATCC 51299 (vancomycin resistant), Escherichia coli ATCC 25922, Listeria monocytogenes ATCC 19111, Mycobacterium tuberculosis H37RvT, Pseudomonas aeruginosa ATCC 27853, Salmonella enterica subsp. arizonae ATCC 13314T, and the methicillin-resistant Staphylococcus aureus subsp. aureus ATCC 33591 (MRSA). The name Streptosporangium minutum is proposed with the type strain M26T (=LMG 28850T =NRRL B-65295T).


Hormonal induction of spermiation, courting behavior and spawning in the southern bell frog, Litoria raniformis.
We trialled the efficacy of various exogenous hormones to induce spermiation, courtship behavior, and spawning in the "endangered" southern bell frog, Litoria raniformis. Intralymphatic administration of Lucrin(®), a synthetic nonapeptide luteinizing hormone releasing hormone (LHRH), was used successfully to induce courting behaviors and ejaculation of spermatozoa in males. Various hormones, including Lucrin(®), another synthetic LHRH analog ([des-Gly(10), D-Ala(6)]-LHRH), human chorionic gonadotropin, progesterone, and a dopamine receptor antagonist failed to promote oviposition and spawning in females. This and earlier studies indicate that in the efficacy of hormonal induction in amphibians varies between taxa, hormones, and genders. The lack of response in females may limit the use of reproduction technology in the southern bell frog and closely related species of Australian bell frogs.


Assessing the Role of Experimental Evidence for Interface Judgment: Licensing of Negative Polarity Items, Scalar Readings, and Focus.
This paper reviews a series of experimental studies that address what we call "interface judgment," which is the complex judgment involving integration from multiple levels of grammatical representation such as the syntax-semantics and prosody-semantics interface. We first discuss the results from the ERP literature connected to NPI licensing in different languages, paying particular attention to the N400 and the P600 as neural correlates of this specific phenomenon and focusing on the study by Xiang et al. (2016). The results of this study show evidence that there are two distinct NPI licensing mechanisms, i.e., licensing and rescuing, in line with Giannakidou (1998, 2006). Then we discuss an acceptability judgment task on Greek NPIs which supports the negativity as a scale hypothesis (Zwarts, 1995, 1996; Giannakidou, 1998). For the semantics-prosody interface judgment, we discuss two types of findings on two different phenomena and languages: (i) the study by Giannakidou and Yoon (2016) on scalar and non-scalar NPIs in Greek and Korean, which serves as the foundation for Chatzikonstantinou's (2016) study of production data showing distinct prosodic properties in emphatic (scalar) and non-emphatic (non-scalar) Greek NPIs; (ii) a (production and perception) study by Etxeberria and Irurtzun (2015) on the prosodic disambiguation of the scalar/non-scalar readings of sentences containing the focus particle "ere" in Basque. The main conclusion of the paper is that experimental methods of the kind discussed in the paper are useful in establishing physical, quantitative correlates of interface judgment.


Disproportional body growth in female estrogen receptor-alpha-inactivated mice.
Estrogens play an important role in the regulation of longitudinal bone growth in man, as demonstrated by recent descriptions of individuals with estrogen insensitivity or aromatase deficiency. Two estrogen receptors, ERalpha and ERbeta, have been cloned. The aim of the present study was to investigate the function of ERalpha in the regulation of body growth and skeletal growth. Adult female mice with inactivated ERalpha (ERalpha-/-) demonstrated an increased body weight compared with wild-type mice (114% of control). However, the length of the appendicular skeleton was decreased in adult ERalpha-/- mice (femur 93% of control). In contrast, the axial skeleton was normal (crown-rump length 98% of control). The decreased growth of the appendicular skeleton was associated with decreased serum levels of IGF-I (77% of control), indicating that the GH/IGF-I axis may be involved in the decreased longitudinal bone growth seen in female ERalpha-/- mice.


Autofocusing Airy beams generated by all-dielectric metasurface for visible light.
Conventional method to generate autofocusing Airy (AFA) beam involves the optical Fourier transform (FT) system, which has a fairly long working distance due to the focal length of FT lens, presence of spatial light modulator (SLM) and auxiliary total reflection mirrors. Here, we propose an extremely compact design to generate high-efficiency AFA beam at visible frequency by using metasurface which is composed of a single layer array of amorphous titanium dioxide (TiO2) elliptical nanofins sitting on the fused-silica substrate. Numerical simulations show that the designed structures are capable of precisely controlling the deflection of Airy beam and tuning the focal length of AFA beam. We further numerically demonstrate that the phase modulation of AFA beam could combine with the concept of vortex light field to produce vortical AFA beam. We anticipate that such device can be useful in the ultra-compact integrated optic system, biomedical nanosurgery and optical trapping applications.


Chronic periodontitis can affect the levels of potential oral cancer salivary mRNA biomarkers.
More than 100 salivary constituents have been found to show levels significantly different in patients with oral squamous cell carcinoma (OSCC) from those found in healthy controls, and therefore have been suggested to be potential salivary biomarkers for OSCC detection. However, many of these potential OSCC salivary biomarkers are also involved in chronic inflammation, and whether the levels of these biomarkers could be affected by the presence of chronic periodontitis was not known. The objective of this pilot study was therefore to measure the levels of seven previously reported potential OSCC salivary mRNA biomarkers in patients with chronic periodontitis and compare them to levels found in patients with OSCC and healthy controls. The seven salivary mRNAs were interleukin (IL)-8, IL-1β, dual specificity phosphatase 1, H3 histone family 3A, ornithine decarboxylase antizyme 1, S100 calcium-binding protein P (S100P) and spermidine/spermine N1-acetyltransferase 1. Unstimulated whole saliva samples were collected from a total of 105 human subjects from the following four study groups: OSCC; CPNS (chronic periodontitis, moderate to severe degree, non-smokers); CPS (chronic periodontitis, moderate to severe degree, smokers); and healthy controls. Levels of each mRNA in patient groups (OSCC or chronic periodontitis) relative to the healthy controls were determined by a pre-amplification reverse transcription-quantitative polymerase chain reaction approach with nested gene-specific primers. Results were recorded and analyzed by the Bio-Rad CFX96 Real-Time System. Mean fold changes between each pair of patient vs. control groups were analyzed by the Mann-Whitney U-test with Bonferroni corrections. Only S100P showed significantly higher levels in patients with OSCC compared to both patients with CPNS (p = 0.003) and CPS (p = 0.007). The difference in S100P levels between patients with OSCC and healthy controls was also marginally significant (p = 0.009). There was no significant difference in the levels of salivary IL-8, IL-1β and dual specificity phosphatase 1 mRNAs between patients with OSCC and patients with CPNS (p = 0.510, 0.058 and 0.078, respectively); no significant difference in levels of salivary ornithine decarboxylase antizyme 1 and spermine N1-acetyltransferase mRNAs between patients with OSCC and patients with CPS (p = 0.318 and 0.764, respectively); and no significant difference in levels of the H3 histone family 3A mRNA between patients with OSCC and either CPS (p = 0.449) or healthy controls (p = 0.107). Salivary S100P mRNA could be a reliable biomarker for OSCC detection, regardless of the presence of chronic periodontitis. The presence of chronic periodontitis could significantly affect the levels of the other six mRNAs, and negatively influence reliability for using them as biomarkers for oral cancer detection.


Outlook of organ transplantation in Iran: time for quality assessment.
Organ transplantation as an undeniable life-saving therapeutic modality fundamentally requires infrastructure, devoted and trained professionals, and positive public attitude to be set up in a well-organized manner at the national level. In addition to sharing achievements and reviewing the increasing trend of transplanted organs in the past 12 years following legislations in Iran, this report raises some concerns from the point of transplantation outcome view.


A spark counter as a control unit of a radio frequency surgery device.
A new approach in radio frequency (rf) electrosurgery, used for tissue treatment, is achieved by using a new process control method. An external control unit allows a commonly available rf-generator to automatically supply the appropriate power for differing tissue types, thus ensuring best cutting quality. The sparks, generated during the scalpel electrode interaction with the tissue, appear statistically distributed. The spark rate depends on various factors and is monotonic with the supplied electrical power. This allows it to be used as the controlled variable in the cutting process. The ac current passing through the tissue is evaluated by an external control unit using an analyzation algorithm to determine the number of sparks. The external unit is comprised of a system, which measures the spark rate, and a subsequent proportional integral (P.I.) controller. The functionality of the control method as well as the electrical circuitry is verified through cuts with different degrees of carbonization and cuts through tissue heterojunctions.


ABHD5 frameshift deletion in Golden Retrievers with ichthyosis.
Ichthyoses are hereditary skin disorders characterized by the formation of scales and defects in the outermost layer of the epidermis. In dogs, at least six different breed-specific ichthyoses including a relatively common PNPLA1-related autosomal recessive ichthyosis in Golden Retrievers are known. In this study, we investigated 14 Golden Retrievers with scales that were not homozygous for the mutant PNPLA1 allele suggesting a genetically distinct new form of ichthyosis. Histopathological examinations showed lamellar, orthokeratotic hyperkeratosis, and mildly hyperplastic epidermis that led to the diagnosis of a nonepidermolytic ichthyosis. Combined linkage and homozygosity mapping in 14 cases and 30 nonaffected family members delimited a critical interval of ∼12.7 Mb on chromosome 23. Whole-genome sequencing of an affected dog revealed a single protein-changing variant within this region that was not present in 795 control genomes. The identified variant is a 14 bp deletion in the ABHD5 gene (c.1006_1019del), leading to a frameshift and altering the last 14 codons p.(Asp336Serfs*6). The genotypes at this variant showed perfect cosegregation with the ichthyosis phenotype in a large family comprising 14 cases and 72 controls. ABHD5 encodes an acyltransferase required for lipid metabolism. In humans, variants in ABHD5 cause Chanarin-Dorfman syndrome, a neutral lipid storage disease with ichthyosis. Our data in dogs together with the knowledge on the effects of ABHD5 variants in humans strongly suggest ABHD5:c.1006_1019del as candidate causative genetic variant for a new canine form of ichthyosis, which we propose to designate as Golden Retriever ichthyosis type 2 (ICH2).


A longitudinal cline characterizes the genetic structure of human populations in the Tibetan plateau.
Indigenous populations of the Tibetan plateau have attracted much attention for their good performance at extreme high altitude. Most genetic studies of Tibetan adaptations have used genetic variation data at the genome scale, while genetic inferences about their demography and population structure are largely based on uniparental markers. To provide genome-wide information on population structure, we analyzed new and published data of 338 individuals from indigenous populations across the plateau in conjunction with worldwide genetic variation data. We found a clear signal of genetic stratification across the east-west axis within Tibetan samples. Samples from more eastern locations tend to have higher genetic affinity with lowland East Asians, which can be explained by more gene flow from lowland East Asia onto the plateau. Our findings corroborate a previous report of admixture signals in Tibetans, which were based on a subset of the samples analyzed here, but add evidence for isolation by distance in a broader geospatial context.


Think again: the role of reappraisal in reducing negative valence bias.
Stimuli such as surprised faces are ambiguous in that they are associated with both positive and negative outcomes. Interestingly, people differ reliably in whether they evaluate these and other ambiguous stimuli as positive or negative, and we have argued that a positive evaluation relies in part on a biasing of the appraisal processes via reappraisal. To further test this idea, we conducted two studies to evaluate whether increasing the cognitive accessibility of reappraisal through a brief emotion regulation task would lead to an increase in positive evaluations of ambiguity. Supporting this prediction, we demonstrated that cuing reappraisal, but not in three other forms of emotion regulation (Study 1a-d; n = 120), increased positive evaluations of ambiguous faces. In a sign of robustness, we also found that the effect of reappraisal generalised from ambiguous faces to ambiguous scenes (Study 2; n = 34). Collectively, these findings suggest that reappraisal may play a key role in determining responses to ambiguous stimuli. We discuss these findings in the context of affective flexibility, and suggest that valence bias (i.e. the tendency to evaluate ambiguity more positively or negatively) represents a novel approach to measuring implicit emotion regulation.


Profanity in media associated with attitudes and behavior regarding profanity use and aggression.
We hypothesized that exposure to profanity in media would be directly related to beliefs and behavior regarding profanity and indirectly to aggressive behavior. We examined these associations among 223 adolescents attending a large Midwestern middle school. Participants completed a number of questionnaires examining their exposure to media, attitudes and behavior regarding profanity, and aggressive behavior. Results revealed a positive association between exposure to profanity in multiple forms of media and beliefs about profanity, profanity use, and engagement in physical and relational aggression. Specifically, attitudes toward profanity use mediated the relationship between exposure to profanity in media and subsequent behavior involving profanity use and aggression. The main hypothesis was confirmed, and implications for the rating industry and research field are discussed.


Maternal mortality following thromboembolism; incidences and prophylaxis strategies.
Thromboembolism is one of the main causes of maternal mortality, which can be prevented in many cases. The present study was designed to investigate the incidence and prophylaxis strategies for maternal mortality following thromboembolism in postnatal. In this case series study, the data of the mortality cases were extracted according to the ethical and security standards of the Ministry of Health of the country and compared with a healthy control group. The thromboembolism risk factors measured and scored using a questionnaire entitled "the evaluation of risk factors for maternal mortality following thromboembolism during pregnancy, labor, or post-partum". The maternal mortality rate was 16 per 100,000 live births. Among 297 mortality cases, 27 (9%) death were due to thromboembolism. The mean gestational age was 32.5 weeks. Dyspnea (88.8%) and tachycardia (18.5%) were found as common clinical manifestations in these patients. Sixteen cases (59.3%) did not get heparin, 6 (22.2%) received single dose and 5 (18.5%) received two doses and more. In these 11 cases, 5 (45%) patients received heparin before surgery, 1 after surgery, and 5 before and after surgery. Twenty cases deceased in the first hours after delivery and the rest after 2 to 12 days. The average score of risk for thromboembolism based on Royal College of Obstetricians & Gynecologist (RCOG) guideline was 4.6. It seems that one of the most important cause of maternal mortality in this study was the lack of recognition of high-risk patients and the lack of prescription for prophylaxis with heparin and this clearly explains the need for accurate screening of high-risk mothers, designing a standard form and the care and treatment of these patients.


TT virus and hepatitis G virus infections in Korean blood donors and patients with chronic liver disease.
To determine the prevalences of TTV and HGV infections among blood donors and patients with chronic liver disease in Korea, to investigate the association of TTV and HGV infections with blood transfusion, and to assess the correlation between TTV and HGV viremia and hepatic damage. A total of 391 serum samples were examined in this study. Samples were obtained from healthy blood donors (n=110), hepatitis B surface antigen (HBsAg)-positive donors (n=112), anti-hepatitis C virus (anti-HCV)-positive donors (n=69), patients with type B chronic liver disease (n=81), and patients with type C chronic liver disease (n=19). TTV DNA was detected using the hemi-nested PCR. HGV RNA was tested using RT-PCR. A history of blood transfusion and serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were also determined. TTV DNA was detected in 8.2 % of healthy blood donors, 16.1 % of HBsAg-positive donors, 20.3 % of anti-HCV-positive donors, 21.0 % of patients with type B chronic liver disease, and 21.1 % of patients with type C chronic liver disease. HGV RNA was detected in 1.8 % of healthy blood donors, 1.8 % of HBsAg-positive donors, 17.4 % of anti-HCV-positive donors, 13.6 % of patients with type B chronic liver disease, and 10.5 % of patients with type C chronic liver disease. The prevalence of TTV and HGV infections in HBV- or HCV-positive donors and patients was significantly higher than in healthy blood donors (P<0.05), except for the detection rate of HGV in HBsAg-positive donors which was the same as for healthy donors. There was a history of transfusion in 66.7 % of TTV DNA-positive patients and 76.9 % of HGV RNA-positive patients (P<0.05). No significant increase in serum ALT and AST was detected in the TTV- or HGV-positive donors and patients. TTV and HGV infections are more frequently found in donors and patients infected with HBV or HCV than in healthy blood donors. However, there is no significant association between TTV or HGV infections and liver injury.


Physical activity and telomere length in early stage breast cancer survivors.
Telomere length (TL) is a biomarker of accumulated cellular damage and human aging. Evidence in healthy populations suggests that TL is impacted by a host of psychosocial and lifestyle factors, including physical activity. This is the first study to evaluate the relationship between self-reported physical activity and telomere length in early stage breast cancer survivors. A cross-sectional sample of 392 postmenopausal women with stage I-III breast cancer at an outpatient oncology clinic of a large university hospital completed questionnaires and provided a blood sample. TL was determined using terminal restriction fragment length analysis of genomic DNA isolated from peripheral blood mononuclear cells. Physical activity was dichotomized into two groups (none versus moderate to vigorous) using the International Physical Activity Questionnaire. Multivariate linear and logistic regression analyses were performed to identify factors associated with mean TL and physical activity. Among participants, 66 (17%) did not participate in any physical activity. In multivariate model adjusted for age, compared to those who participated in moderate to vigorous physical activity, women who participated in no physical activity had significantly shorter TL (adjusted coefficient β=-0.22; 95% confidence interval (CI), -0.41 to -0.03; P=.03). Non-white race, lower education and depressive symptoms were associated with lack of self-reported physical activity (P<0.05 for all) but not TL. Lack of physical activity is associated with shortened TL, warranting prospective investigation of the potential role of physical activity on cellular aging in breast cancer survivors.


Total intravenous anesthesia, spinal anesthesia or combined sciatic-femoral nerve block for outpatient knee arthroscopy.
The aim of this study was to compare efficacy, efficiency and surgeon's satisfaction of total intravenous anesthesia with propofol and remifentanil with those of spinal or peripheral nerve blocks for outpatient knee arthroscopy. One hundred and twenty patients undergoing elective outpatient knee arthroscopy were randomly allocated to receive total intravenous anesthesia with propofol and remifentanil (40), combined sciatic-femoral nerve block (40), or spinal anesthesia (40). Preparation times, surgeon's satisfaction, and discharge times with the 3 anesthesia techniques were measured. Anesthesia-related costs were also compared based on costs of drugs, disposable materials, and anesthesia and nurse staff. Preparation time was 13 min (8-22 min) with general anesthesia, 15 min (5-30 min) with spinal anesthesia and 15 min (5-25 min) with sciatic-femoral blocks (p=0.006). Surgeon's satisfaction was similar in the 3 groups, but 17 patients receiving peripheral nerve block (42%) and 12 receiving spinal anesthesia (30%) by-passed the postanesthesia care unit after surgery as compared with only 2 general anesthesia patients (5%) (p=0.01). Discharge from the postanesthesia care unit required 5 min (5-20 min) after peripheral block as compared with 15 min (5-25 min) with spinal and 15 min (5-80 min) with general anesthesia (p=0.005); however, stay in the Day-Surgery Unit was shorter after general anesthesia [170 (100-400) min] than peripheral [265 (110-485) min] or spinal blocks [230 (95-800) min] (p=0.026). Urinary retention was reported in 3 spinal patients only (8%) (p=0.03). Regional anesthesia techniques reduce the rate of admission and the duration of stay in the postanesthesia care unit as compared with general anesthesia. Peripheral rather than spinal nerve blocks should be preferred to minimise the risk for urinary retention.


Report of a case of Nezelof syndrome.
In this report, we present a 5 months old male baby, who suffered from watery diarrhea since 4 days old. From then on, he had been admitted 3 times in 3 different hospitals but the symptoms still bothered him off and on. During the days of hospitalization, sepsis with positive blood culture of Klebsiella was noted. The patient expired at 5 months of age. The T cell count was 20% active T was 0. Delayed hypersensitivity skin tests including Candida (10 X), PHA (10 micrograms), PHA (1 microgram), SK/SD (50 units) were negative. The granulocyte function study showed normal. Immunoglobulin analysis revealed IgG: 1320 mg%, IgA: 120 mg%, IgM: 100 mg%. Agenesis of thymus, failure of lymphoid differentiation and abnormal lymphoid architecture with absence of germinal centers were noted at autopsy. Combined immunodeficiency with normal immunoglobulins (Nezelof syndrome) is a disease of primary immunodeficiency characterized by recurrent infections, failure to thrive, lymphopenia, diminished lymphoid tissue, abnormal structure or agenesis of the thymus, and presence of normal or increased levels of one or more of the major immunoglobulin classes, but with impaired antibody synthesis. Since its original description by Nezelof and associates in 1964, it has been reported on the subsequent occasion. In this report, we present our one experience and review the clinical and laboratory data in 33 reported cases.


Group sequential two-stage preference designs.
The two-stage preference design (TSPD) enables inference for treatment efficacy while allowing for incorporation of patient preference to treatment. It can provide unbiased estimates for selection and preference effects, where a selection effect occurs when patients who prefer one treatment respond differently than those who prefer another, and a preference effect is the difference in response caused by an interaction between the patient's preference and the actual treatment they receive. One potential barrier to adopting TSPD in practice, however, is the relatively large sample size required to estimate selection and preference effects with sufficient power. To address this concern, we propose a group sequential two-stage preference design (GS-TSPD), which combines TSPD with sequential monitoring for early stopping. In the GS-TSPD, pre-planned sequential monitoring allows investigators to conduct repeated hypothesis tests on accumulated data prior to full enrollment to assess study eligibility for early trial termination without inflating type I error rates. Thus, the procedure allows investigators to terminate the study when there is sufficient evidence of treatment, selection, or preference effects during an interim analysis, thereby reducing the design resource in expectation. To formalize such a procedure, we verify the independent increments assumption for testing the selection and preference effects and apply group sequential stopping boundaries from the approximate sequential density functions. Simulations are then conducted to investigate the operating characteristics of our proposed GS-TSPD compared to the traditional TSPD. We demonstrate the applicability of the design using a study of Hepatitis C treatment modality.


Physiological characteristics of the biocontrol yeast Pichia anomala J121.
The yeast Pichia anomala J121 prevents mold spoilage and enhances preservation of moist grain in malfunctioning storage systems. Development of P. anomala J121 as a biocontrol agent requires in-depth knowledge about its physiology. P. anomala J121 grew under strictly anaerobic conditions, at temperatures between 3 degrees C and 37 degrees C, at pH values between 2.0 and 12.4, and at a water activity of 0.92 (NaCl) and 0.85 (glycerol). It could assimilate a wide range of C- and N-sources and produce killer toxin. A selective medium containing starch, nitrate, acetic acid, and chloramphenicol was developed for P. anomala. P. anomala was equally sensitive as Candida albicans to common antifungal compounds. Growth ability at a range of environmental conditions contributes to the competitive ability of the biocontrol yeast P. anomala J121.


The conflict between cheetahs and humans on Namibian farmland elucidated by stable isotope diet analysis.
Large areas of Namibia are covered by farmland, which is also used by game and predator species. Because it can cause conflicts with farmers when predators, such as cheetahs (Acinonyx jubatus), hunt livestock, we assessed whether livestock constitutes a significant part of the cheetah diet by analysing the stable isotope composition of blood and tissue samples of cheetahs and their potential prey species. According to isotopic similarities, we defined three isotopic categories of potential prey: members of a C4 food web with high δ15N values (gemsbok, cattle, springhare and guinea fowl) and those with low δ15N values (hartebeest, warthog), and members of a C3 food web, namely browsers (eland, kudu, springbok, steenbok and scrub hare). We quantified the trophic discrimination of heavy isotopes in cheetah muscle in 9 captive individuals and measured an enrichment for 15N (3.2‰) but not for 13C in relation to food. We captured 53 free-ranging cheetahs of which 23 were members of groups. Cheetahs of the same group were isotopically distinct from members of other groups, indicating that group members shared their prey. Solitary males (n = 21) and males in a bachelor groups (n = 11) fed mostly on hartebeest and warthogs, followed by browsers in case of solitary males, and by grazers with high δ15N values in case of bachelor groups. Female cheetahs (n = 9) predominantly fed on browsers and used also hartebeest and warthogs. Mixing models suggested that the isotopic prey category that included cattle was only important, if at all, for males living in bachelor groups. Stable isotope analysis of fur, muscle, red blood cells and blood plasma in 9 free-ranging cheetahs identified most individuals as isotopic specialists, focussing on isotopically distinct prey categories as their food.


Cell-based angiopoietin-1 gene therapy for acute lung injury.
The acute respiratory distress syndrome is a significant cause of morbidity and mortality in critically ill patients. Angiopoietin-1 (Ang-1), a ligand for the endothelial Tie2 receptor, is an endothelial survival and vascular stabilization factor that reduces endothelial permeability and inhibits leukocyte-endothelium interactions. We hypothesized that Ang-1 counteracts vascular inflammation and pulmonary vascular leak in experimental acute lung injury. We used cell-based gene therapy in a rat model of ALI. Transgenic mice overexpressing Ang-1 or deficient in the Tie2 receptor were also studied to better elucidate the mechanisms of protection. The present report provides data that support a strong protective role for the Ang-1/Tie2 system in two experimental models of LPS-induced acute lung injury. In a rat model, cell-based Ang-1 gene transfer improved morphological, biochemical, and molecular indices of lung injury and inflammation. These findings were confirmed in a gain-of-function conditional, targeted transgenic mouse model, in which Ang-1 reduced endothelial cell activation and the expression of adhesion molecules, associated with a marked improvement in airspace inflammation and intraalveolar septal thickening. Moreover, heterozygous Tie2-deficient mice demonstrated enhanced evidence of lung injury and increased early mortality. These results support a critical role for the Ang-1/Tie2 axis in modulating the pulmonary vascular response to lung injury and suggest that Ang-1 therapy may represent a potential new strategy for the treatment and/or prevention of acute respiratory distress syndrome in critically ill patients.


De novo proteins from designed combinatorial libraries.
Combinatorial libraries of de novo amino acid sequences can provide a rich source of diversity for the discovery of novel proteins with interesting and important activities. Randomly generated sequences, however, rarely fold into well-ordered proteinlike structures. To enhance the quality of a library, features of rational design must be used to focus sequence diversity into those regions of sequence space that are most likely to yield folded structures. This review describes how focused libraries can be constructed by designing the binary pattern of polar and nonpolar amino acids to favor proteins that contain abundant secondary structure, while simultaneously burying hydrophobic side chains and exposing hydrophilic side chains to solvent. The "binary code" for protein design was used to construct several libraries of de novo proteins, including both alpha-helical and beta-sheet structures. The recently determined solution structure of a binary patterned four-helix bundle is well ordered, thereby demonstrating that sequences that have neither been selected by evolution (in vivo or in vitro) nor designed by computer can form nativelike proteins. Examples are presented demonstrating how binary patterned libraries have successfully produced well-ordered structures, cofactor binding, catalytic activity, self-assembled monolayers, amyloid-like nanofibrils, and protein-based biomaterials.


Deep Learning Empowered Fresnel-based Lensless Fluorescence Microscopy.
Miniaturized fluorescence microscopy has revolutionized the way neuroscientists study the brain in-vivo. Recent developments in computational lensless imaging promise a next generation of miniaturized microscopes in lensless fluorescence microscopy. We developed a microscope prototype using an optimized Fresnel amplitude mask. While many lensless imaging modalities have reported excellent performance using Deep Learning (DL) approaches, DL application in fluorescence imaging has been left untouched. We generated a computational dataset based on experimental system calibration to evaluate DL capabilities on biological cell morphologies. We show that our DL-assisted microscope can provide high-quality imaging with a structural similarity index of 89%. The least absolute error was decreased by 63% using the DL-assisted method compared with the classical models. The state-of-the-art performance of this prototype enhances the expected potential of amplitude masks in lensless microscopy applications, which are critical for robust in-vivo flat microscopy with engineered image sensors.Clinical Relevance- This study aids in advancing miniaturized fluorescence microscopy, which greatly impacts long-term brain circuit and disease studies in freely moving animal models.


Carbon nanotubes as photoacoustic molecular imaging agents in living mice.
Photoacoustic imaging of living subjects offers higher spatial resolution and allows deeper tissues to be imaged compared with most optical imaging techniques. As many diseases do not exhibit a natural photoacoustic contrast, especially in their early stages, it is necessary to administer a photoacoustic contrast agent. A number of contrast agents for photoacoustic imaging have been suggested previously, but most were not shown to target a diseased site in living subjects. Here we show that single-walled carbon nanotubes conjugated with cyclic Arg-Gly-Asp (RGD) peptides can be used as a contrast agent for photoacoustic imaging of tumours. Intravenous administration of these targeted nanotubes to mice bearing tumours showed eight times greater photoacoustic signal in the tumour than mice injected with non-targeted nanotubes. These results were verified ex vivo using Raman microscopy. Photoacoustic imaging of targeted single-walled carbon nanotubes may contribute to non-invasive cancer imaging and monitoring of nanotherapeutics in living subjects.


Electrophoretic mobility and molecular distribution studies of poly(amidoamine) dendrimers of defined charges.
Generation 5 ethylenediamine (EDA)-cored poly(amidoamine) (PAMAM) dendrimers (E5, E denotes the EDA core and 5 the generation number) with different degrees of acetylation and carboxylation were synthesized and used as a model system to investigate the effect of charge and the influence of dendrimer surface modifications on electrophoretic mobility (EM) and molecular distribution. The surface-modified dendrimers were characterized by size-exclusion chromatography, 1H NMR, MALDI-TOF-MS, PAGE, and CE. The focus of our study was to determine how EM changes as a function of particle charge and molecular mass, and how the molecular distribution changes due to surface modifications. We demonstrate that partially modified dendrimers have much broader migration peaks than those of fully surface functionalized or unmodified E5 dendrimers due to variations in the substitution of individual dendrimer surfaces. EM decreased nonlinearly with increases in surface acetylation for both PAMAM acetamides and PAMAM succinamic acids, indicating a complex migration activity in CE separations that is not solely due to charge/mass ratio changes. These studies provide new insights into dendrimer properties under an electric field, as well as into the characterization of dendrimer-based materials being developed for medical applications.


Impact of salinity on the population dynamics of microorganisms in a membrane bioreactor treating produced water.
Biomass characteristics may change subject to a salinity increase when treating high strength wastewater. In this study, the impact of salinity in a membrane bioreactor (MBR) was investigated for the treatment of produced water (PW). MBR was operated as a pre-treatment prior to nanofiltration (NF) and reverse osmosis (RO). Mixed PW, that was originated from different oil, gas, and oil-gas wells, were subjected to pre-treatment for 146 days including three different operational phases: Low-salinity (~10 mS/cm), gradual increased salinity (10-40 mS/cm) and high salinity (~40 mS/cm). The results of this study showed that microorganisms could adapt using real wastewater and treat PW up to a certain level of the salinity. At high salinity levels of PW, the floc structure started to be disrupted and membrane fouling was accelerated. PCR-DGGE analysis confirmed the changes in microbial communities' composition in relation with high salinity. The results of the final treatment experiments presented that NF and RO treatment produced high-quality effluents that could be suitable for reuse.


Peptide binding to metal oxide nanoparticles.
Magnetic metal oxide nanoparticles demonstrate great applicability in several fields such as biotechnology, medicine and catalysis. A stable, magnetic and low-cost material, nanoscale magnetite, is an interesting adsorbent for protein purification. Downstream processing can account for up to 80% of the total production costs in biotechnological production. As such, the development of new innovative separation methods can be regarded as highly profitable. While short peptide sequences can be used as specific affinity tags for functionalised adsorber surfaces, they need expensive affinity ligands on the particle surface for adsorption. In order to identify peptide tags for several non-functionalised inorganic surfaces, different binding conditions to iron oxide nanoparticles are evaluated. Therefore, magnetite nanoparticles in a range of 5-20 nm were synthesised with a co-precipitation method. Zeta potential measurements indicated an amphiphilic surface with an isoelectric point in the neutral pH region. Glutamic acid-based homo-peptides were used as affinity peptides for the magnetite nanoparticles. We demonstrate a dependence of the binding affinity of the peptides on pH and buffer ions in two different experimental set-ups. The nature of surface coordination for glutamic acid-based peptides can be demonstrated with different spectroscopic approaches such as infrared spectroscopy (IR), Raman spectroscopy and circular dichroism spectroscopy (CD). We want to emphasise the importance of physicochemical properties such as surface energy, polarity, morphology and charge. These parameters, which are dependent on the environmental conditions, play a crucial role in peptide interactions with iron oxide surfaces. The understanding of the adsorption of simple biomolecules on nanoscale metal oxide surfaces also represents the key to the even more complex interactions of proteins at the bio-nano interface. From the identification of interaction patterns and an understanding of the adsorption of these peptides, the up-scaling to tagged model proteins facilitates the possibility of an industrial magnetic separation process and might therefore reduce time and costs in purification processes.


Proliferation, migration and differentiation of neuronal progenitor cells in the adult mouse subventricular zone surgically separated from its olfactory bulb.
The subventricular zone of the adult mammalian forebrain contains progenitor cells that, by migrating along a restricted pathway called the 'rostral migratory stream' (RMS), add new neurons to the olfactory bulb throughout life. To determine the influence of the olfactory bulb on the development of these progenitor cells, we performed lesions that interrupt this pathway and separate the olfactory bulb from the rest of the forebrain. By labelling cells born at several survival times after the lesions with the thymidine analogue bromodeoxyuridine (BrdU), we found that disconnection from the bulb influences the rate of BrdU incorporation by the progenitor cells. The number of labelled cells in lesioned mice was almost half that found in control mice. In the disconnected migratory pathway, the number of neurons expressing calretinin was increased indicating that neuronal differentiation was enhanced: newly born neurons occurred within and around the RMS, most of them expressed calretinin and left the pathway starting about 2 weeks after the lesion. Thereafter, these neurons preserving their phenotype, spread for long distances, and accumulated ectopically in dorsal regions of the anterior olfactory nucleus and the frontal cortex. Finally, transplantation of adult subventricular cells into the lesioned pathway showed that the lesion neither prevents neuronal migration nor alters its direction. Thus, although the olfactory bulb appears to regulate the pace of the developmental processes, its disconnection does not prevent the proliferation, migration and phenotypic acquisition of newly generated bulbar interneurons that, since they cannot reach their terminal domains, populate some precise regions of the lesioned adult forebrain.


Direct Comparison of Cardiac Myosin-Binding Protein C With Cardiac Troponins for the Early Diagnosis of Acute Myocardial Infarction.
Cardiac myosin-binding protein C (cMyC) is a cardiac-restricted protein that is more abundant than cardiac troponins (cTn) and is released more rapidly after acute myocardial infarction (AMI). We evaluated cMyC as an adjunct or alternative to cTn in the early diagnosis of AMI. Unselected patients (N=1954) presenting to the emergency department with symptoms suggestive of AMI, concentrations of cMyC, and high-sensitivity (hs) and standard-sensitivity cTn were measured at presentation. The final diagnosis of AMI was independently adjudicated using all available clinical and biochemical information without knowledge of cMyC. The prognostic end point was long-term mortality. Final diagnosis was AMI in 340 patients (17%). Concentrations of cMyC at presentation were significantly higher in those with versus without AMI (median, 237 ng/L versus 13 ng/L, P<0.001). Discriminatory power for AMI, as quantified by the area under the receiver-operating characteristic curve (AUC), was comparable for cMyC (AUC, 0.924), hs-cTnT (AUC, 0.927), and hs-cTnI (AUC, 0.922) and superior to cTnI measured by a contemporary sensitivity assay (AUC, 0.909). The combination of cMyC with hs-cTnT or standard-sensitivity cTnI (but not hs-cTnI) led to an increase in AUC to 0.931 (P<0.0001) and 0.926 (P=0.003), respectively. Use of cMyC more accurately classified patients with a single blood test into rule-out or rule-in categories: Net Reclassification Improvement +0.149 versus hs-cTnT, +0.235 versus hs-cTnI (P<0.001). In early presenters (chest pain <3 h), the improvement in rule-in/rule-out classification with cMyC was larger compared with hs-cTnT (Net Reclassification Improvement +0.256) and hs-cTnI (Net Reclassification Improvement +0.308; both P<0.001). Comparing the C statistics, cMyC was superior to hs-cTnI and standard sensitivity cTnI (P<0.05 for both) and similar to hs-cTnT at predicting death at 3 years. cMyC at presentation provides discriminatory power comparable to hs-cTnT and hs-cTnI in the diagnosis of AMI and may perform favorably in patients presenting early after symptom onset. URL: https://www.clinicaltrials.gov. Unique identifier: NCT00470587.


Understanding and practice: a 7-year follow-up study on implementation of a cardiac rehabilitation program.
The authors explore the perspectives of managers, health care professionals, and patients on a comprehensive cardiac rehabilitation program. Using qualitative methodology, they compare and analyze results from individual interviews and two conceptual modeling seminars held 7 years apart. Professionals and managers understood their own tasks in a professional-centered way that did not include the client's perspective. Patients believed they were not seen in their whole context. Initially, health care organization was fragmented, lacking clear leadership, coordination, and communication between levels of care. However, lack of common understandings of structure, process, and outcome in cardiac rehabilitation services hampered the implementation of program changes.


Cryptic genetic variation and paraphyly in ravens.
Widespread species that are morphologically uniform may be likely to harbour cryptic genetic variation. Common ravens (Corvus corax) have an extensive range covering nearly the entire Northern Hemisphere, but show little discrete phenotypic variation. We obtained tissue samples from throughout much of this range and collected mitochondrial sequence and nuclear microsatellite data. Our study revealed a deep genetic break between ravens from the western United States and ravens from throughout the rest of the world. These two groups, the 'California clade' and the 'Holarctic clade' are well supported and over 4% divergent in mitochondrial coding sequence. Microsatellites also reveal significant differentiation between these two groups. Ravens from Minnesota, Maine and Alaska are more similar to ravens from Asia and Europe than they are to ravens from California. The two clades come in contact over a huge area of the western United States, with mixtures of the two mitochondrial groups present in Washington, Idaho and California. In addition, the restricted range Chihuahuan raven (Corvus cryptoleucus) of the south-west United States and Mexico is genetically nested within the paraphyletic common raven. Our findings suggest that the common raven may have formerly consisted of two allopatric groups that may be in the process of remerging.


The management of spinal deformities in Duchenne muscular dystrophy. A new concept of spinal bracing.
A clinical study of 62 patients with Duchenne Muscular Dystrophy has suggested 2 pathways in the pathogenesis of the spinal deformities. In the first pathway, the patients develop an initial kyphosis due to the loss of paravertebral muscle thus placing the intrinsic ligamentous spine in an unstable position. The spine then develops a lateral curve accentuated by pelvic obliquity. In the final form, axial rotation of the spine and pelvis occurs to produce a severe deformity. In the second pathway the intrinsic spine assumes the more stable position of hyperextension with a level pelvis. This position remains relatively unchanged during the period of wheelchair confinement even though muscle deterioration progresses. A significant increase in the severity of the deformity occurs with age in those patients with unstable spines. None of the other factors examined affected the severity of the lateral curvature. A spinal brace has been designed to keep the spine extended and the pelvis level while the patients sits in his wheelchair. The purpose is to encourage the inexorable progression of muscular deficiencies to follow the second pathway. Various modifications of wheelchairs also may help to maintain spinal stability.


Association between Dietary Factors and Constipation in Adults Living in Luxembourg and Taking Part in the ORISCAV-LUX 2 Survey.
Constipation, a disorder of bowel movements, is among the most frequent gastrointestinal complaints in Western countries. Dietary constituents such as inadequate fiber intake have been related to constipation, but discrepancies exist in the findings regarding dietary factors. This study investigated the association between dietary patterns and bowel movements in adults living in Luxembourg. Data from 1431 participants from ORISCAV-LUX 2 (a cross-sectional survey) who completed a 174-item food frequency questionnaire (FFQ) were analyzed. A questionnaire-based constipation score was assessed by a validated scoring system. Confounders such as physical activity and serum/urine indicators were assessed. Women had higher constipation scores than men (p < 0.001). In food group-based regression models, a negative association was found between higher constipation score and intake of grains (Beta = -0.62, 95%CI: -1.18, -0.05) and lipid-rich foods (Beta = -0.84, 95%CI: -1.55, -0.13), while a positive association was found for sugary products (Beta = 0.54, 95%CI: 0.11, 0.97) (p < 0.05). In a nutrient-based regression model, a positive association was found between constipation score and total energy (Beta = 5.24, 95%CI: 0.37, 10.11) as well as sodium intake (Beta = 2.04, 95%CI: 0.21, 3.87), and a negative one was found for total fats (Beta = -4.17, 95%CI: -7.46, -0.89) and starch (Beta = -2.91, 95%CI: -4.47, -1.36) (p < 0.05). Interestingly, neither fruits and vegetables or dietary fiber were significantly associated with constipation. Thus, grains, lipid-rich foods, total fats and starch were associated with a lower constipation score, while sugary products, sodium, and higher energy intake were correlated with higher constipation.


Effects of different Helicobacter pylori culture filtrates on growth of gastric epithelial cells.
To study the effects of different Helicobacter pylori (H pylori) culture filtrates on growth of gastric epithelial cells. Broth culture filtrates of H pylori were prepared. Gastric epithelial cells were treated with the filtrates, and cell growth was determined by growth curve and flow cytometry. DNA damage of gastric epithelial cells was measured by single-cell microgel electrophoresis. Gastric epithelial cells proliferated actively when treated by CagA-gene-positive broth culture filtrates, and colony formation reached 40%. The number of cells in S phase increased compared to controls. Comet assay showed 41.2% comet cells in GES-1 cells treated with CagA-positive filtrates (P < 0.05). CagA-positive filtrates enhance the changes in morphology and growth characteristics of human gastric epithelial tumor cells. DNA damage maybe one of the mechanisms involved in the growth changes.


Synergistic signals in the mechanism of antigen-induced exocytosis in 2H3 cells: evidence for an unidentified signal required for histamine release.
The aim of this study was to determine whether the increase in cytosolic free Ca2+ concentration ([Ca2+]i) in response to antigen (aggregated ovalbumin) on IgE-primed 2H3 cells was sufficient to account for exocytosis. When the [Ca2+]i responses to antigen and the Ca2+ ionophore A23187 were compared, A23187 was much less effective at releasing histamine at equivalent [Ca2+]i increases, and little or no stimulated histamine release occurred with A23187 concentrations that matched the [Ca2+]i response to antigen concentrations that stimulated maximal histamine release. The [Ca2+]i response to antigen is not, therefore, sufficient to account for exocytosis, although extracellular Ca2+ is necessary to initiate both the [Ca2+]i response and histamine release: the antigen must generate an additional, unidentified, signal that is required for exocytosis. To determine whether this signal was the activation of protein kinase C, the effects of the phorbol ester 12-0-tetradecanoyl phorbol 13-acetate (TPA) on the responses to antigen were examined. TPA blocked the antigen-induced [Ca2+]i response and the release of inositol phosphates but had little effect on histamine release and did not stimulate exocytosis by itself. The unidentified signal from the antigen is therefore distinct from the activation of protein kinase C and is generated independently of the [Ca2+]i response or the release of inositol phosphates. Taken together with other data that imply that there is very little activation of protein kinase C by antigen when the rate of histamine release is maximal, it is concluded that the normal exocytotic response to antigen requires the synergistic action of the [Ca2+]i signal together with an unidentified signal that is not mediated by protein kinase C.


Identification of novel immune infiltration-related biomarkers of sepsis based on bioinformatics analysis.
This study aimed to analyze the gene expression characteristics of sepsis and search for potential biomarkers involved in the pathogenesis. The data on sepsis were obtained from the GEO database according to the keyword "sepsis". The CIBERSORT algorithm was applied to determine the immune cell. WGCNA package were applied to build a weighted gene network. Then, a topological overlap matrix was created and dynamic hybrid cutting was applied to categorizing the genes with identical expression patterns. Component analysis of each module was implemented according to module eigengenes. In order to detect the important modules, the connections among the immune infiltration of Mφ and the modules were computed by Pearson's tests. PPI network was made using the STRING database and cytoHubba was applied to find hub genes. A total of 760 sepsis samples as well as 42 healthy control samples were involved. A total of 14 gene modules were generated. Thereinto, the correlations of the yellow (includes 168 hub genes) and blue (includes 526 hub genes) modules with Mφ0 were 0.39 and -0.42, while with Mφ1 were 0.3 and -0.4. 916 up-regulated and 454 down-regulated DEGs were found in the sepsis group. 451 intersected genes were obtained after the intersecting of DEGs and the hub genes from blue and yellow modules. Subsequent GO enrichment analysis suggested that 451 overlapping genes were enriched in "T cell activation", "lymphocyte differentiation" and "T cell differentiation" for biological process. Besides, KEGG enrichment analysis showed that "Human T-cell leukemia virus 1 infection" and "Th17 cell differentiation" were the most enriched pathways. In PPI network, UTP6, RRS1, RRP1B, DDX18, and DDX24 were identified as hub genes. ROC analysis showed the AUC values of these genes were all greater than 0.95. UTP6, RRS1, RRP1B, DDX18, and DDX24 participate in the pathogenetic process of sepsis through regulating the activation and differentiation of lymphocytes. Besides, these five genes could be used for diagnosing sepsis.


Biology-based modeling to analyze uranium toxicity data on Daphnia magna in a multigeneration study.
Recent studies have investigated chronic toxicity of waterborne depleted uranium on the life cycle and physiology of Daphnia magna. In particular, a reduction in food assimilation was observed. Our aims here were to examine whether this reduction could fully account for observed effects on both growth and reproduction, for three successive generations, and to investigate through microscope analyses whether this reduction resulted from direct damage to the intestinal epithelium. We analyzed data obtained by exposing Daphnia magna to uranium over three successive generations. We used energy-based models, which are both able to fit simultaneously growth and reproduction and are biologically relevant. Two possible modes of action were compared - decrease in food assimilation rate and increase in maintenance costs. In our models, effects were related either to internal concentration or to exposure concentration. The model that fitted the data best represented a decrease in food assimilation related to exposure concentration. Furthermore, observations of consequent histological damage to the intestinal epithelium, together with uranium precipitates in the epithelial cells, supported the assumption that uranium has direct effects on the digestive tract. We were able to model the data in all generations and showed that sensitivity increased from one generation to the next, in particular through a significant increase of the intensity of effect, once the threshold for appearance of effects was exceeded.


Genetic and chemical correction of cholesterol accumulation and impaired autophagy in hepatic and neural cells derived from Niemann-Pick Type C patient-specific iPS cells.
Niemann-Pick type C (NPC) disease is a fatal inherited lipid storage disorder causing severe neurodegeneration and liver dysfunction with only limited treatment options for patients. Loss of NPC1 function causes defects in cholesterol metabolism and has recently been implicated in deregulation of autophagy. Here, we report the generation of isogenic pairs of NPC patient-specific induced pluripotent stem cells (iPSCs) using transcription activator-like effector nucleases (TALENs). We observed decreased cell viability, cholesterol accumulation, and dysfunctional autophagic flux in NPC1-deficient human hepatic and neural cells. Genetic correction of a disease-causing mutation rescued these defects and directly linked NPC1 protein function to impaired cholesterol metabolism and autophagy. Screening for autophagy-inducing compounds in disease-affected human cells showed cell type specificity. Carbamazepine was found to be cytoprotective and effective in restoring the autophagy defects in both NPC1-deficient hepatic and neuronal cells and therefore may be a promising treatment option with overall benefit for NPC disease.


Response to Hypoxia and the Ensuing Dysregulation of Inflammation Impacts Mycobacterium tuberculosis Pathogenicity.
Rationale: Different Mycobacterium tuberculosis (Mtb) strains exhibit variable degrees of virulence in humans and animal models. Differing stress response strategies used by different strains of Mtb could influence virulence. Objectives: We compared the virulence of two strains of Mtb with use in animal model research: CDC1551 and Erdman. Methods: Rhesus macaques, which develop human-like tuberculosis attributes and pathology, were infected with a high dose of either strain via aerosol, and virulence was compared by bacterial burden and pathology. Measurements and Main Results: Infection with Erdman resulted in significantly shorter times to euthanasia and higher bacterial burdens and greater systemic inflammation and lung pathology relative to those infected with CDC1551. Macaques infected with Erdman also exhibited significantly higher early inflammatory myeloid cell influx to the lung, greater macrophage and T cell activity, and higher expression of lung remodeling (extracellular matrix) genes, consistent with greater pathology. Expression of NOTCH4 (neurogenic locus notch homolog 4) signaling, which is induced in response to hypoxia and promotes undifferentiated cellular state, was also higher in Erdman-infected lungs. The granulomas generated by Erdman, and not CDC1551, infection appeared to have larger regions of necrosis, which is strongly associated with hypoxia. To better understand the mechanisms of differential hypoxia induction by these strains, we subjected both to hypoxia in vitro. Erdman induced higher concentrations of DosR regulon relative to CDC1551. The DosR regulon is the global regulator of response to hypoxia in Mtb and critical for its persistence in granulomas. Conclusions: Our results show that the response to hypoxia is a critical mediator of virulence determination in Mtb, with potential impacts on bacillary persistence, reactivation, and efficiency of therapeutics.


Aridity drove the evolution of extreme embolism resistance and the radiation of conifer genus Callitris.
Xylem vulnerability to embolism is emerging as a major factor in drought-induced tree mortality events across the globe. However, we lack understanding of how and to what extent climate has shaped vascular properties or functions. We investigated the evolution of xylem hydraulic function and diversification patterns in Australia's most successful gymnosperm clade, Callitris, the world's most drought-resistant conifers. For all 23 species in this group, we measured embolism resistance (P50 ), xylem specific hydraulic conductivity (Ks ), wood density, and tracheary element size from natural populations. We investigated whether hydraulic traits variation linked with climate and the diversification of this clade using a time-calibrated phylogeny. Embolism resistance varied widely across the Callitris clade (P50 : -3.8 to -18.8 MPa), and was significantly related to water scarcity, as was tracheid diameter. We found no evidence of a safety-efficiency tradeoff; Ks and wood density were not related to rainfall. Callitris diversification coincides with the onset of aridity in Australia since the early Oligocene. Our results highlight the evolutionary lability of xylem traits with climate, and the leading role of aridity in the diversification of conifers. The uncoupling of safety from other xylem functions allowed Callitris to evolve extreme embolism resistance and diversify into xeric environments.


Excuses to continue smoking: the role of disengagement beliefs in smoking cessation.
The aim of the present study was to investigate the role of disengagement beliefs in smoking cessation. The association of disengagement beliefs with forward transition through the transtheoretical stages of change and self-reported quitting were examined, with and without adjusting for processes of change. A longitudinal survey was conducted among Dutch smokers, resulting in 367 respondents who completed two questionnaires, one at baseline and one at 8 months follow-up. Disengagement beliefs, intention to quit, actual quitting, and the processes of change were assessed. The findings showed that disengagement beliefs were negatively associated with forward stage transition and actual quitting. Processes of change only partly mediated these associations. Adhering to disengagement beliefs seems to be an inhibitor of progression towards smoking cessation and actual quitting, also after adjustment for processes of change.


Chronic ethanol consumption in rats produces opioid antinociceptive tolerance through inhibition of mu opioid receptor endocytosis.
It is well known that the mu-opioid receptor (MOR) plays an important role in the rewarding properties of ethanol. However, it is less clear how chronic ethanol consumption affects MOR signaling. Here, we demonstrate that rats with prolonged voluntary ethanol consumption develop antinociceptive tolerance to opioids. Signaling through the MOR is controlled at many levels, including via the process of endocytosis. Importantly, agonists at the MOR that promote receptor endocytosis, such as the endogenous peptides enkephalin and β-endorphin, show a reduced propensity to promote antinociceptive tolerance than do agonists, like morphine, which do not promote receptor endocytosis. These observations led us to examine whether chronic ethanol consumption produced opioid tolerance by interfering with MOR endocytosis. Indeed, here we show that chronic ethanol consumption inhibits the endocytosis of MOR in response to opioid peptide. This loss of endocytosis was accompanied by a dramatic decrease in G protein coupled receptor kinase 2 (GRK2) protein levels after chronic drinking, suggesting that loss of this component of the trafficking machinery could be a mechanism by which endocytosis is lost. We also found that MOR coupling to G-protein was decreased in ethanol-drinking rats, providing a functional explanation for loss of opioid antinociception. Together, these results suggest that chronic ethanol drinking alters the ability of MOR to endocytose in response to opioid peptides, and consequently, promotes tolerance to the effects of opioids.


Sequence-dependent cargo recognition by SNX-BARs mediates retromer-independent transport of CI-MPR.
Endosomal recycling of transmembrane proteins requires sequence-dependent recognition of motifs present within their intracellular cytosolic domains. In this study, we have reexamined the role of retromer in the sequence-dependent endosome-to-trans-Golgi network (TGN) transport of the cation-independent mannose 6-phosphate receptor (CI-MPR). Although the knockdown or knockout of retromer does not perturb CI-MPR transport, the targeting of the retromer-linked sorting nexin (SNX)-Bin, Amphiphysin, and Rvs (BAR) proteins leads to a pronounced defect in CI-MPR endosome-to-TGN transport. The retromer-linked SNX-BAR proteins comprise heterodimeric combinations of SNX1 or SNX2 with SNX5 or SNX6 and serve to regulate the biogenesis of tubular endosomal sorting profiles. We establish that SNX5 and SNX6 associate with the CI-MPR through recognition of a specific WLM endosome-to-TGN sorting motif. From validating the CI-MPR dependency of SNX1/2-SNX5/6 tubular profile formation, we provide a mechanism for coupling sequence-dependent cargo recognition with the biogenesis of tubular profiles required for endosome-to-TGN transport. Therefore, the data presented in this study reappraise retromer's role in CI-MPR transport.


Chlorine inactivation of Salmonella Kentucky isolated from chicken carcasses: evaluation of strain variation.
The current study was undertaken to evaluate chlorine resistance among strains of Salmonella Kentucky isolated from chicken carcasses. Selected strains (n = 8) were exposed to 30 ppm of chlorine in 10% buffered peptone water (pH 7.4) for 0 to 10 min at 4°C and 150 rpm. The initial level (mean ± SD) of Salmonella Kentucky was 6.18 ± 0.09 log CFU/ml and did not differ (P > 0.05) among strains. A two-way analysis of variance indicated that the level of Salmonella Kentucky in chlorinated water was affected (P < 0.05) by a time by strain interaction. Differences among strains increased as a function of chlorine exposure time. After 10 min of chlorine exposure, the most resistant strain (SK145) was 5.63 ± 0.54 log CFU/ml, whereas the least resistant strain (SK275) was 3.07 ± 0.29 log CFU/ml. Significant differences in chlorine resistance were observed for most strain comparisons. Death of Salmonella Kentucky was nonlinear over time and fitted well to a power law model with a shape parameter of 0.34 (concave upward). Time (minutes) for a 1-log reduction of Salmonella Kentucky differed (P < 0.05) among strains: >10 min for SK145, 6.0 min for SK254, 1.5 min for SK179, and 0.3 to 0.65 min for other strains. Results of this study indicate that strain is an important variable to include in models that predict changes in levels of Salmonella Kentucky in chlorinated water.


Evaluation of intracranial meningioma resection with a surgical aspirator in dogs: 17 cases (1996-2004).
To determine results of intracranial meningioma resection by use of a surgical aspirator and assess prognostic factors associated with intracranial meningiomas in dogs. Retrospective case series. 17 dogs. Medical records of dogs that underwent resection of an intracranial meningioma by use of a surgical aspirator were reviewed. Information pertaining to signalment, imaging findings, clinical signs, duration of clinical signs, preoperative treatment, location of the tumor, results of histologic assessment, outcome, and necropsy results was obtained from the medical record. Clients and referring veterinarians were contacted via telephone for information on recurrence of clinical signs and postoperative survival time. 16 dogs were > 7 years of age, and all 17 dogs had seizures before surgery. The most commonly affected breed was the Golden Retriever, represented by 6 of the 17 dogs. Median survival time was 1,254 days. Of the data collected, only histologic subtype of the tumor was prognostic. Analysis of survival times according to histologic tumor subtypes indicated that the order from most brief to longest was as follows: anaplastic, 0 days; fibroblastic, 10 days; psammomatous, > 313 days; meningothelial, > 523 days; and transitional, 1,254 days. Use of a surgical aspirator to resect intracranial meningiomas in dogs was associated with longer survival times than those achieved with traditional surgery alone or traditional surgery combined with radiation therapy. Dogs with meningothelial, psammomatous, or transitional intracranial meningioma subtypes appeared to have a better prognosis than dogs with other subtypes of meningioma.


The effect of extraarticular varus and valgus deformity on total knee arthroplasty.
Because of trauma, metabolic bone disease, congenital deformity, or prior osteotomy, an extraarticular deformity may be present in patients requiring total knee arthroplasty. If the extraarticular deformity is not corrected extraarticularly, it must be corrected by compensatory distal femoral or proximal tibial wedge resection to produce overall limb alignment. Because such a wedge resection between the proximal and distal attachments of the collateral ligaments will produce asymmetrical ligament length, complex instabilities may result. This article, through overlay templates and trigonometric analysis, evaluates all the issues confronting the surgeon deciding whether to pursue intraarticular or extraarticular correction. The conclusions are as follows: (1) the closer a deformity is to the knee, the greater its importance, (2) femoral deformities are more difficult to correct intraarticularly than tibial deformities because femoral compensatory wedge resection produces instability only in extension, and (3) intraarticular correction of varus deformities produces lateral instability that is usually better tolerated than medial instability, and some extraarticular deformities are best treated by extraarticular correct, independent, or total knee arthroplasty.