Automated Myocardial Infarction Screening Using Morphology-Based Electrocardiogram Biomarkers.
Ischemic heart disease (IHD), a critical and dreadful cardiovascular disease, is a leading cause of death globally. The steady progress of IHD leads to an irreversible condition called myocardial infarction (MI). The detection of MI can be done by observing the altered electrocardiogram (ECG) characteristics. Often, automated ECG analysis is preferred in place of visual inspection to reduce time and ensure reliable detection even when the recording quality is not very good. This paper presents an automated approach to classify recent MI, past MI, and normal sinus rhythm (NSR) classes based on the morphological features of the ECG. In clinical practice, a standard 12-lead ECG setup is typically employed to identify MI. However, acquiring a 12-lead ECG is not always convenient. Hence, in this study, we have explored the possibility of using a minimal number of ECG leads by deriving the augmented limb leads using leads I and II. A well-known and widely used ensemble machine learning tool, the random forest (RF) classifier is trained using features extracted from the derived augmented limb leads and their combinations. An RF classifier built using features extracted from all limb leads has outperformed classifiers built on combinations of them with five-fold cross-validation training accuracy of 97.9 (±0.008) % and testing accuracy of 98 %.Clinical relevance- As high sensitivity is reported in identifying recent MI and past MI classes, the proposed approach is suitable for preventative healthcare applications since it is less likely that subjects with recent or past MI will be misclassified. Due to its low computational complexity, better interpretability, and comparable performance to the state-of-the-art results, the proposed approach can be employed in clinical and cardiac health screening applications. It also has the potential to be employed in remote monitoring with mobile and wearable devices because it is built on features extracted from only lead I and II ECG recordings.


Flocs surface functionality assessment of sonicated activated sludge in relation with physico-chemical properties.
Flocs surface functional groups evolutions due to an ultrasonic treatment were investigated in respect with the mechanisms involved during sonication. Activated sludge surface functional groups changes were studied after treatment of a sample at different ultrasonic specific energies. Sludge functionality was qualitatively assessed by recording the infrared (FT-IR) spectra of centrifugation pellets. Potentiometric titration coupled with proton surface complexation modeling was used to assess the nature and quantity of ionizable functional groups present at the floc surface and in the aqueous phase. These evolutions were linked to changes of both mixed liquor biochemical composition (TSS, VSS, COD, proteins, humic like substances, polysaccharides) and physical properties (floc size and settleability). Observations carried out showed that activated sludge flocs were essentially mechanically disintegrated by ultrasonic waves: the nature of chemical bonds observed by FT-IR did not shift after ultrasonic treatment. Moreover, the total number of ionizable functional groups measured by potentiometric titrations remained constant during sonication. However, due to the solubilization of organic components induced by cavitation process, the corresponding ionizable functional groups (carboxyl, hydroxyl, amine) were transferred from particulate to soluble fraction. Moreover, due to the variable amount of proteins, humic like substances and polysaccharides solubilised, the relative contributions of carboxyl, hydroxyl and amine groups varied at floc surface.


A carrier-mediated transport for folate in basolateral membrane vesicles of rat small intestine.
The mechanism of exit of folate from the enterocyte, i.e. transport across the basolateral membrane, is not known. In this study we examined, using basolateral membrane vesicles, the transport of folic acid across the basolateral membrane of rat intestine. Uptake of folic acid by these vesicles represents transport of the substrate into the intravesicular compartment and not binding to the membrane surface. The rate of folic acid transport was linear for the first 1 min of incubation but decreased thereafter, reaching equilibrium after 5 min of incubation. The transport of folic acid was: (1) saturable as a function of concentration with an apparent Km of 0.6 +/- 0.17 microM and Vmax. of 1.01 +/- 0.11 pmol/30 s per mg of protein; (2) inhibited in a competitive manner by the structural analogues 5-methyltetrahydrofolate and methotrexate (Ki = 2 and 1.4 microM, respectively); (4) electroneutral; (5) Na+-independent; (6) sensitive to the effect of the anion exchange inhibitor 4,4'-di-isothiocyanatostilbene-2,2'-disulphonic acid (DIDS). These data indicate the existence of a carrier-mediated transport system for folic acid in rat intestinal basolateral membrane and demonstrate that the transport process is electroneutral, Na+-independent and sensitive to the effect of anion exchange inhibition.


The efficacy of the novel dual PI3-kinase/mTOR inhibitor NVP-BEZ235 compared with rapamycin in renal cell carcinoma.
Inhibitors of TORC1 have been shown to be active in patients with metastatic renal cell carcinoma (RCC). As the phosphatidylinositol 3-kinase (PI3K) pathway activates numerous other kinases, transcription factors, and proteins associated with cell growth and survival besides mammalian target of rapamycin (mTOR), disruption of this pathway upstream of mTOR may be more effective than inhibition of TORC1 alone. To investigate this possibility, the dual PI3K/mTOR inhibitor NVP-BEZ235 was compared with rapamycin in RCC cell lines and xenografts generated from 786-O and A498 cells. Treatment of RCC cell lines with NVP-BEZ235 in vitro resulted in the nuclear translocation of p27, greater reduction in tumor cell proliferation, and more complete suppression of Akt, Mnk-1, eIF4E, and 4EBP-1 phosphorylation and cyclin D1 and hypoxia-inducible factor 2alpha (HIF2alpha) expression than that achieved with rapamycin. The reduction of HIF2alpha levels correlated with reduced HIF activity as determined by luciferase assay. NVP-BEZ235 induced growth arrest in both the 786-O and A498 xenografts that was associated with inhibition of Akt and S6 phosphorylation as well as the induction of apoptosis and reduction in markers of tumor cell proliferation. In contrast, rapamycin induced only minimal growth retardation. Dual inhibition of PI3K/mTOR with NVP-BEZ235 induced growth arrest in RCC cell lines both in vitro and in vivo more effectively than inhibition of TORC1 alone. These results provide the rationale for the clinical assessment of agents such as NVP-BEZ235 in patients with advanced RCC.


Stage-variations of anandamide hydrolase activity in the mouse uterus during the natural oestrus cycle.
Recent studies have demonstrated that the endogenous cannabinoids are important modulators of fertility in mammals. In particular, a role of the endocannabinoid system in early stages of embryo development, oviductal transport of embryos, pregnancy maintenance and labour has been demonstrated in rodents and/or in humans. In the present paper, we report the analysis of FAAH activity and protein content in the mouse uterus as a function of the natural oestrus cycle stages. Variations of FAAH activity are discussed in relationship to changes in sex steroid levels and to the possible action of AEA on remodelling of uterine tissues.


The effect of CreA in glucose and xylose catabolism in Aspergillus nidulans.
The catabolism of glucose and xylose was studied in a wild type and creA deleted (carbon catabolite de-repressed) strain of Aspergillus nidulans. Both strains were cultivated in bioreactors with either glucose or xylose as the sole carbon source, or in the presence of both sugars. In the cultivations on single carbon sources, it was demonstrated that xylose acted as a carbon catabolite repressor (xylose cultivations), while the enzymes in the xylose utilisation pathway were also subject to repression in the presence of glucose (glucose cultivations). In the wild type strain growing on the sugar mixture, glucose repression of xylose utilisation was observed; with xylose utilisation occurring only after glucose was depleted. This phenomenon was not seen in the creA deleted strain, where glucose and xylose were catabolised simultaneously. Measurement of key metabolites and the activities of key enzymes in the xylose utilisation pathway revealed that xylose metabolism was occurring in the creA deleted strain, even at high glucose concentrations. Conversely, in the wild type strain, activities of the key enzymes for xylose metabolism increased only when the effects of glucose repression had been relieved. Xylose was both a repressor and an inducer of xylanases at the same time. The creA mutation seemed to have pleiotropic effects on carbohydratases and carbon catabolism.


NLRP3 promotes autophagy of urate crystals phagocytized by human osteoblasts.
Monosodium urate (MSU) microcrystals present in bone tissues of chronic gout can be ingested by nonprofessional phagocytes like osteoblasts (OBs) that express NLRP3 (nucleotide-binding domain and leucine-rich repeat region containing family of receptor protein 3). MSU is known to activate NLRP3 inflammasomes in professional phagocytes. We have identified a new role for NLRP3 coupled to autophagy in MSU-stimulated human OBs. Normal human OBs cultured in vitro were investigated for their capacity for phagocytosis of MSU microcrystals by using confocal microscopy. Subsequent mineralization and matrix metalloproteinase activity were evaluated, whereas regulatory events of phagocytosis were deciphered by using signaling inhibitors, phosphokinase arrays, and small interfering RNAs. Statistics were carried out by using paired or unpaired t tests, and the one-way ANOVA, followed by multiple comparison test. Most of the OBs internalized MSU in vacuoles. This process depends on signaling via PI3K, protein kinase C (PKC), and spleen tyrosine kinase (Syk), but is independent of Src kinases. Simultaneously, MSU decreases phosphorylation of the protein kinases TOR (target of rapamycin) and p70S6K. MSU activates the cleavage of microtubule-associated protein light chain 3 (LC3)-I into LC3-II, and MSU microcrystals are coated with GFP-tagged LC3. However, MSU-stimulated autophagy in OBs absolutely requires the phagocytosis process. We find that MSU upregulates NLRP3, which positively controls the formation of MSU-autophagosomes in OBs. MSU does not increase death and late apoptosis of OBs, but reduces their proliferation in parallel to decreasing their competence for mineralization and to increasing their matrix metalloproteinase activity. MSU microcrystals, found locally encrusted in the bone matrix of chronic gout, activate phagocytosis and NLRP3-dependent autophagy in OBs, but remain intact in permanent autophagosomes while deregulating OB functions.


RNF213 rare variants in an ethnically diverse population with Moyamoya disease.
Moyamoya disease (MMD) is a rare, genetically heterogeneous cerebrovascular disease resulting from occlusion of the distal internal carotid arteries. A variant in the Ring Finger 213 gene (RNF213), altering arginine at position 4810 (p.R4810K), is associated with MMD in Asian populations. However, there are a lack of data on the role of RNF213 in patients with MMD of additional ethnicities and diasporic Asian populations. We investigate the contribution of RNF213 alterations to MMD in an ethnically diverse population based in the United States. We initially sequenced RNF213 exons 43, 44, and 45 (encoding the eponymous RING finger domain) and exon 60 (encoding p.R4810K) in 86 ethnically diverse patients with MMD. Comprehensive exome sequencing data from 24 additional patients with MMD was then analyzed to identify RNF213 variants globally. Segregation of variants with MMD and other vascular diseases was assessed in families. RNF213 p.R4810K was identified in 56% (9/16) of patients with MMD of Asian descent and not in 94 patients of non-Asian descent. 3.6% (4/110) of patients had variants in the exons encoding the RING finger domain. Seven additional variants were identified in 29% (7/24) of patients with MMD who underwent exome sequencing. Segregation analysis supported an association with MMD for 2 variants and a lack of association with disease for 1 variant. These results confirm that alterations in RNF213 predispose patients of diverse ethnicities to MMD, and that the p.R4810K variant predisposes individuals of Asian descent in the United States to MMD.


An angular motion of a conserved four-helix bundle facilitates alternating access transport in the TtNapA and EcNhaA transporters.
There is ongoing debate regarding the mechanism through which cation/proton antiporters (CPAs), like Thermus thermophilus NapA (TtNapA) and Escherichia coli NapA (EcNhaA), alternate between their outward- and inward-facing conformations in the membrane. CPAs comprise two domains, and it is unclear whether the transition is driven by their rocking-bundle or elevator motion with respect to each other. Here we address this question using metadynamics simulations of TtNapA, where we bias conformational sampling along two axes characterizing the two proposed mechanisms: angular and translational motions, respectively. By applying the bias potential for the two axes simultaneously, as well as to the angular, but not the translational, axis alone, we manage to reproduce each of the two known states of TtNapA when starting from the opposite state, in support of the rocking-bundle mechanism as the driver of conformational change. Next, starting from the inward-facing conformation of EcNhaA, we sample what could be its long-sought-after outward-facing conformation and verify it using cross-linking experiments.


Microbiological and biochemical properties of canestrato pugliese hard cheese supplemented with bifidobacteria.
Canestrato Pugliese cheeses from ewe milk were produced according to a traditional protocol and by adding 7.0 log10 cfu of fresh cells per gram of Bifidobacterium bifidum Bb02, Bifidobacterium longum Bb46, or both species. The traditional technology was modified slightly to favor the survival of probiotic microorganisms. After 56 d of ripening, the survival of B. bifidum Bb02 and B. longum Bb46 was 6.0 and 5.0 log10 cfu/g, respectively. After 19 d cheeses contained ca. 7.0 log10 cfu/g of bifidobacteria. Compared to traditional cheese, the addition of bifidobacteria seemed to support the growth and survival of mesophilic lactobacilli and Streptococcus thermophilus, used as starter, during ripening. No significant differences were observed in the main chemical composition, and only a slightly higher concentration of acetic acid was found in cheeses with bifidobacteria added. On the contrary, alpha- and beta-galactosidase activities were markedly more pronounced in the presence of bifidobacteria, especially with B. bifidum Bb02. In contrast with traditional cheese, the lactose was completely hydrolyzed in cheeses made with bifidobacteria. Urea-PAGE electrophoresis of the pH 4.6-soluble and pH 4.6-insoluble N fractions did not show appreciable variations. Only the reversed-phase-HPLC analysis of the pH 4.6-soluble N showed a slightly more complex profile in the presence of bifidobacteria. This finding was in agreement with the higher value of the pH 4.6-soluble N/total N ratio and with the more pronounced amino-, imino-, and dipeptidase activities found in all the cheeses with the bifidobacteria added, especially B. bifidum Bb02. No differences were found in the free amino acid and free fatty acid contents. The amino acids glutamic acid, valine, proline, leucine, and lysine and the fatty acids butyric, caproic, capric, and oleic acids were found at the highest concentrations. The sensory evaluation did not show significant differences, and Canestrato Pugliese cheeses were characterized by small and uniformly distributed eyes, were pale yellow, had an elastic consistency and a Pecorino-like smell, were very salty, and tended to be moderately piquant.


Dynamic stereochemical rearrangements in chiral organometallic complexes.
Organometallic complexes have long been known to display a wide variety of dynamic stereochemical processes. Classic examples of such processes include the exchange of axial and equatorial environments in trigonal bipyramidal complexes, such as Fe(CO)(5), and the migration of the metal moiety round the periphery of the cyclopentadiene ring in eta(1)-bound Cp complexes. The systematic study of fluxional processes is of interest because it can not only help provide a detailed, quantitative 'picture' of the bonding between the metals and ligands involved, but it can also help to rationalise chemical reactivity patterns. The introduction of chirality into organometallic complexes, usually in the form of a non-racemic chiral ligand, has led to an explosion in the importance such species, particularly with regard to their applications in organic functional group transformations. The presence of a chiral centre can also provide an excellent spectroscopic handle on the complex in question, enabling both novel fluxional processes to be observed and new light to be shed on old (unresolved) problems. In this critical review (101 references) the literature on metal-centred fluxional rearrangements in chiral transition and main group organometallic complexes is reviewed, complementing the recent review by Faller (see reference 8).


Worldwide Network for Blood and Marrow Transplantation (WBMT) recommendations for establishing a hematopoietic stem cell transplantation program in countries with limited resources (Part II): Clinical, technical and socio-economic considerations.
The development of hematopoietic stem cell transplantation (HSCT) programs can face significant challenges in most developing countries because such endeavors must compete with other government health care priorities, including the delivery of basic services. While this is may be a limiting factor, these countries should prioritize development of the needed expertise to offer state of the art treatments including transplantation, by providing financial, technological, legal, ethical and other needed support. This would prove beneficial in providing successful programs customized to the needs of their population, and potentially provide long-term cost-savings by circumventing the need for their citizens to seek care abroad. Costs of establishing HSCT program and the costs of the HSCT procedure itself can be substantial barriers in developing countries. Additionally, socioeconomic factors intrinsic to specific countries can influence access to HSCT, patient eligibility for HSCT and timely utilization of HSCT center capabilities. This report describes recommendations from the Worldwide Network for Blood and Marrow Transplantation (WBMT) for establishing HSCT programs with a specific focus on developing countries, and identifies challenges and opportunities for providing this specialized procedure in the resource constrained setting.


Tumors of the appendix: I. Neoplastic and nonneoplastic mucoceles.
We reviewed the clinical data and pathologic material on 46 patients with lesions originally interpreted as appendiceal mucocele or mucinous adenocarcinoma from two medical centers. The mucoceles, characterized by accumulation of mucus in a dilated lumen, could be divided into three distinct entities: nonneoplastic retention mucoceles (eight cases), cystadenomas (26 cases), and cystadenocarcinomas (eight cases). There were four mucinous adenocarcinomas. Retention mucoceles are associated with little morbidity and are cured by appendectomy. Cystadenomas and cystadenocarcinomas mimic the behavior of ovarian neoplasms of borderline malignant potential. Some of these lesions were associated with pseudomyxoma peritonei, but did not have systemic metastases. Mucinous adenocarcinomas without mucocele mimic the behavior of ordinary large bowel carcinoma.


Bacillus oryzaecorticis sp. nov., a moderately halophilic bacterium isolated from rice husks.
A Gram-stain-positive, aerobic, endospore-forming, moderately halophilic rod, designated strain R1(T), was isolated from rice husks and subjected to a taxonomic study using a polyphasic approach. Strain R1(T) produced spherical or ellipsoidal endospores at a subterminal position in swollen sporangia, and was catalase- and oxidase-positive. The isolate grew optimally at 37 °C and pH 6.0-7.0, and could grow in the presence of up to 9% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain R1(T) belongs to the genus Bacillus. The closest relatives of strain R1(T) were Bacillus subtilis subsp. subtilis NCIB 3610(T), Bacillus aquimaris TF-12(T), and Bacillus marisflavi TF-11(T), with 16S rRNA gene sequence similarities of 96.0%, 98.4%, and 98.7%, respectively. DNA-DNA relatedness values between the isolate and the reference strains were ≤42±3%. The predominant menaquinones were MK-5 (50%) and MK-7 (50%). The major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylethanolamine. The major cellular fatty acids were iso-C(15 : 0) (48.6%) and anteiso-C(15 : 0) (20.6%), and the cell-wall diamino acid was meso-diaminopimelic acid. On the basis of 16S rRNA gene sequence analyses and chemotaxonomic and phenotypic characteristics, it is concluded that strain R1(T) represents a novel species of the genus Bacillus, for which we propose the name Bacillus oryzaecorticis sp. nov. The type strain is R1(T) ( = KACC 17217(T) = KCCM 90231(T) = JCM 19602(T)).


Functional and structural changes throughout the auditory system following congenital and early-onset deafness: implications for hearing restoration.
The absence of auditory input, particularly during development, causes widespread changes in the structure and function of the auditory system, extending from peripheral structures into auditory cortex. In humans, the consequences of these changes are far-reaching and often include detriments to language acquisition, and associated psychosocial issues. Much of what is currently known about the nature of deafness-related changes to auditory structures comes from studies of congenitally deaf or early-deafened animal models. Fortunately, the mammalian auditory system shows a high degree of preservation among species, allowing for generalization from these models to the human auditory system. This review begins with a comparison of common methods used to obtain deaf animal models, highlighting the specific advantages and anatomical consequences of each. Some consideration is also given to the effectiveness of methods used to measure hearing loss during and following deafening procedures. The structural and functional consequences of congenital and early-onset deafness have been examined across a variety of mammals. This review attempts to summarize these changes, which often involve alteration of hair cells and supporting cells in the cochleae, and anatomical and physiological changes that extend through subcortical structures and into cortex. The nature of these changes is discussed, and the impacts to neural processing are addressed. Finally, long-term changes in cortical structures are discussed, with a focus on the presence or absence of cross-modal plasticity. In addition to being of interest to our understanding of multisensory processing, these changes also have important implications for the use of assistive devices such as cochlear implants.


Serious Adverse Events and Their Impact on Functional Outcome in Acute Ischemic Stroke in the WAKE-UP Trial.
During the first days and weeks after an acute ischemic stroke, patients are prone to complications that can influence further treatment, recovery, and functional outcome. In clinical trials, severe complications are recorded as serious adverse events (SAE). We analyzed the effect of SAE on functional outcome and predictors of SAE in the randomized controlled WAKE-UP trial (Efficacy and Safety of MRI-Based Thrombolysis in Wake-Up Stroke). We performed a post hoc analysis of WAKE-UP, a multicenter, randomized, placebo-controlled clinical trial of magnetic resonance imaging-guided intravenous thrombolysis with alteplase in patients with acute ischemic stroke and unknown time of onset. Functional outcome was assessed by the modified Rankin Scale 90 days after the stroke. SAE were reported to a central safety desk and recorded and categorized by organ system using Medical Dictionary for Regulatory Activities terminology. We used logistic regression analysis to determine the effect of SAE on functional outcome and linear multiple regression analysis to identify baseline predictors of SAE. Among 503 patients randomized, 199 SAE were reported for n=110 (22%) patients. Of those patients who did suffer a SAE, 20 (10%) had a fatal outcome. Patients suffering from at least one SAE had a lower odds of reaching a favorable outcome (modified Rankin Scale score of 0-1) at 90 days (adjusted odds ratio, 0.36 [95% CI, 0.21-0.61], P<0.001). Higher age (P=0.04) and male sex (P=0.01) were predictors for the occurrence of SAE. SAEs were observed in about one in 5 patients, were more frequent in elderly and male patients and were associated with worse functional outcome. These results may help to assess the risk of SAE in future stroke trials and create awareness for severe complications after stroke in clinical practice. Registration: URL: https://www.clinicaltrials.gov; Unique identifier: NCT01525290. URL: https://eudract.ema.europa.eu; Unique identifier: 2011-005906-32.


Quantitative multichannel NC-AFM data analysis of graphene growth on SiC(0001).
Noncontact atomic force microscopy provides access to several complementary signals, such as topography, damping, and contact potential. The traditional presentation of such data sets in adjacent figures or in colour-coded pseudo-three-dimensional plots gives only a qualitative impression. We introduce two-dimensional histograms for the representation of multichannel NC-AFM data sets in a quantitative fashion. Presentation and analysis are exemplified for topography and contact-potential data for graphene grown epitaxially on 6H-SiC(0001), as recorded by Kelvin probe force microscopy in ultrahigh vacuum. Sample preparations by thermal decomposition in ultrahigh vacuum and in an argon atmosphere are compared and the respective growth mechanisms discussed.


The development of the skeletal system in children and the influence of muscular strength.
In a study on 14 healthy children aged from 6 to 13 years, the volumetric spongiosa bone density (SBD), cortical bone density (CBD), bone cross-sectional area (BCSA), cortical area (CA), and bone strength index (BSI) at the distal radius were analyzed using peripheral quantitative computer tomography (pQCT Bone Scanner XCT-900, Stratec, Pforzheim, Germany). BSI values were calculated on the basis of the moment of resistance and cortical density using system integrated software. These parameters were correlated with grip strength measurements obtained using a grip dynamometer. There were age-dependent increases in BCSA (r = 0.68), CA (r = 0.78), BSI (r = 0.79). SBD and CBD, however, did not show an increase with age. Grip strength correlates strongly with the parameters of bone geometry BCSA (r = 0.8), CA (r = 0.86), and BSI (r = 0.9) at p < 0.01. SBD and CBD showed no significant correlation to grip strength. The results of this study suggest that an increase in cortical thickness and cross-sectional area represent the most important adaptation mechanisms of the growing bones in response to biomechanical usage. These parameters represent the most important factors of bone strength. Volumetric bone density (material characteristic) of spongiosa and cortical bone is an age-independent parameter which does not change significantly with increasing muscular strength in healthy subjects. These relationships must be considered when selecting parameters to be used as success criteria in studies on the influence of physical exercise on bone status.


Markers of acute rejection and graft acceptance in liver transplantation.
The evaluation of the immunosuppression state in liver transplanted patients is crucial for a correct post-transplant management and a major step towards the personalisation of the immunosuppressive therapy. However, current immunological monitoring after liver transplantation relies mainly on clinical judgment and on immunosuppressive drug levels, without a proper assessment of the real suppression of the immunological system. Various markers have been studied in an attempt to identify a specific indicator of graft rejection and graft acceptance after liver transplantation. Considering acute rejection, the most studied markers are pro-inflammatory and immunoregulatory cytokines and other proteins related to inflammation. However there is considerable overlap with other conditions, and only few of them have been validated. Standard liver tests cannot be used as markers of graft rejection due to their low sensitivity and specificity and the weak correlation with the severity of histopathological findings. Several studies have been performed to identify biomarkers of tolerance in liver transplanted patients. Most of them are based on the analysis of peripheral blood samples and on the use of transcriptional profiling techniques. Amongst these, NK cell-related molecules seem to be the most valid marker of graft acceptance, whereas the role CD4+CD25+Foxp3+ T cells has still to be properly defined.


Esophagitis Dissecans Superficialis in Children.
Esophagitis dissecans superficialis (EDS) is a desquamative disorder of the superficial esophageal epithelium with variable clinical characteristics. Endoscopically, there is an appearance of superficial peeling of sheets of epithelium. Histologically there is 2-toned epithelium with coagulative necrosis of the superficial epithelium. Currently, there is paucity of data regarding this condition in children. A 10-year retrospective search of the pathology information system was performed for cases with a pathologic diagnosis of EDS in a tertiary care pediatric center. Demographic data, clinical history, endoscopic findings, and histopathologic reports were reviewed. Thirteen patients (9 girls; ages 3-18 years), were identified with histologic findings of EDS. Esophageal food impaction, dysphagia, vomiting, and abdominal pain were the most common presenting symptoms. Sixty-nine percentage of the patients had underlying comorbidities and 76% were on at least 1 medication chronically. Eosinophilic esophagitis (23%), inflammatory bowel disease (23%), and gastroesophageal reflux disease (GERD) (15%) were the most common associated diagnoses. Of the 13 patients, 5 had repeat endoscopies showing complete resolution of EDS with no complications. EDS is an under-recognized entity that endoscopists should be familiar with. In our series, the most prevalent associations were with food impaction and eosinophilic esophagitis (EoE). Contact injury and/or inflammation may precede the development of EDS. Pediatric EDS appears to be an incidental finding without significant morbidity or mortality.


Proteomic and Biochemical Analyses of the Mechanism of Tolerance in Mutant Soybean Responding to Flooding Stress.
To investigate the mechanism of flooding tolerance of soybean, flooding-tolerant mutants derived from gamma-ray irradiated soybean were crossed with parent cultivar Enrei for removal of other factors besides the genes related to flooding tolerance in primary generated mutant soybean. Although the growth of the wild type was significantly suppressed by flooding compared with the non-flooding condition, that of the mutant lines was better than that of the wild type even if it was treated with flooding. A two-day-old mutant line was subjected to flooding for 2 days and proteins were analyzed using a gel-free/label-free proteomic technique. Oppositely changed proteins in abundance between the wild type and mutant line under flooding stress were associated in endoplasmic reticulum according to gene-ontology categorization. Immunoblot analysis confirmed that calnexin accumulation increased in both the wild type and mutant line; however, calreticulin accumulated in only the mutant line under flooding stress. Furthermore, although glycoproteins in the wild type decreased by flooding compared with the non-flooding condition, those in the mutant line increased even if it was under flooding stress. Alcohol dehydrogenase accumulated in the wild type and mutant line; however, this enzyme activity significantly increased and mildly increased in the wild type and mutant line, respectively, under flooding stress compared with the non-flooding condition. Cell death increased and decreased in the wild type and mutant line, respectively, by flooding stress. These results suggest that the regulation of cell death through the fermentation system and glycoprotein folding might be an important factor for the acquisition of flooding tolerance in mutant soybean.


[Impact of calcium channel antagonists for estrogen action on the endometrial carcinoma HEC-1A cells].
To study the effects of nifedipine and mibefradil on the cell proliferation, apoptosis, migration on the HEC-1A in vitro and also study the mRNA and protein expression levels of the calcium channel alpha1D (Cav1.3) and calcium channel alpha1G (Cav3.1) to discuss the effects of the calcium antagonists on the mechanisms of the endometrial carcinoma. (1) Add 10 µmol/L nifedipine and mibefradil at 15 minutes before adding 10 µmol/L 17β-estradiol (17β-E(2)) and 100 µmol/L b-estradiol-6-(O-carboxymethyl)oxime (E(2)-BSA) to the HEC-1A in different time including 0, 5, 15, 30, 60, 120 minutes. Then the changes of mRNA and protein were detected by reverse transcripiton (RT)-PCR and western-blot. (2) Add 1.25, 2.5, 5, 10, 20, 40, 80, 100 µmol/L nifedipine and mibefradil to the HEC-1A at 24, 48, 96 hours to detect the cell proliferation by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) method. (3) Add 10 µmol/L nifedipine and mibefradil to the HEC-1A, then detect the apoptosis at 0 minute, 30 minutes, 1 hour, 6 hours, 24 hours and migration in vitro at 36 hours with transwell methods. (1) After the pretreated effect of the nifedipine before 17β-E(2), the mRNA express of Cav1.3 genes was lowest at 15 minutes, and returned to the control level after 30 minutes. The protein level didn't change very much in 30 minutes, but rose after 60 minutes. The Cav3.1 genes mRNA express was lowest at 5 minutes, rose at 30 minutes and returned to the 0 minute level gradually. (2) After the pretreated effect of the nifedipine before E(2)-BSA, the Cav1.3 genes mRNA was lowest at 5 minutes and returned at 15 minutes. The protein level rose gradually in 15 minutes but reduced after 15 minutes. The Cav3.1 mRNA and protein level were reduced at every time point. (3) After the pretreated effect of the mibefradil before 17β-E(2), there was no change of mRNA expression of Cav1.3 genes. The protein level rose at 15 and 60 minutes, there was no change in any other time. The Cav3.1 genes mRNA were gradually reduced and the protein level rose at 15 minutes, and there was no change in any other times. (4) After the pretreated effect of the mibefradil before E(2)-BSA, the mRNA and protein of Cav1.3 levels were reduced after 15 minutes. There was no mRNA expression of Cav3.1, while the protein level was lowest at 15 minutes. (5) Nifedipine and mibefradil affected HEC-1A proliferation depended on the different concentration and interval time points. There was significant difference than those in control group (P < 0.05). (6) There were statistical differences in apoptosis rate after adding nifedipine (P < 0.05), while rose at mibefradil treated the same time (24 hours: 8.41 ± 0.07, 0 minute: 3.74 ± 0.18; P < 0.05). (7) The numbers of stained cells after both nifedipine and mibefradil treated reduced more than control group. (1) Nifedipine and mibefradil could inhibit both the effect of the estrogen on the L-type and T-type calcium channel in short time, meanwhile the mibefradil effects last long time. (2) The inhibited effect of the mibefradil on the proliferation, apoptosis and migration of HEC-1A cells in vitro is more significant than that by nifedidipine.


Mediterranean diet and cognitive function: the SUN project.
Our aim was to evaluate the association between adherence to the Mediterranean Diet (MedDiet) and cognitive function in 823 participants (62 ± 6 years at baseline) from a Spanish prospective cohort (SUN project). A validated 136-item food frequency questionnaire was used to assess the adherence to the MedDiet at baseline. The 10-point (0 to 9) MedDiet Score was used to categorize adherence to MedDiet. Cognitive function was assessed twice at follow-up with a mean follow-up time between exposure and outcome assessment of 6 and 8y using the Telephone Interview of Cognitive Status-modified (TICS-m, range 0 to 54 points). ANCOVA models were used to assess the association between adherence to the MedDiet and cognitive decline. In the multivariable-adjusted analysis of 2-year changes, a higher cognitive decline was observed among participants with low or moderate baseline adherence to the MedDiet than among those with better adherence (adjusted difference = -0.56 points in TICS-m, 95% CI = -0.99 to -0.13). A higher adherence to the MedDiet might be associated with better cognitive function. However, observed differences were of small magnitude and further studies are needed to confirm this finding.


Fluoride exposure and kidney and liver function among adolescents in the United States: NHANES, 2013-2016.
Hepato- and nephrotoxicity of fluoride have been demonstrated in animals, but few studies have examined potential effects in humans. This population-based study examines the relationship between chronic low-level fluoride exposure and kidney and liver function among United States (U.S.) adolescents. This study aimed to evaluate whether greater fluoride exposure is associated with altered kidney and liver parameters among U.S. youth. This cross-sectional study utilized data from the National Health and Nutrition Examination Survey (2013-2016). We analyzed data from 1983 and 1742 adolescents who had plasma and water fluoride measures respectively and did not have kidney disease. Fluoride was measured in plasma and household tap water. Kidney parameters included estimated glomerular filtration rate (calculated by the original Schwartz formula), serum uric acid, and the urinary albumin to creatinine ratio. Liver parameters were assessed in serum and included alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, blood urea nitrogen, gamma-glutamyl transferase, and albumin. Survey-weighted linear regression examined relationships between fluoride exposure and kidney and liver parameters after covariate adjustment. A Holm-Bonferroni correction accounted for multiple comparisons. The average age of adolescents was 15.4 years. Median water and plasma fluoride concentrations were 0.48 mg/L and 0.33 μmol/L respectively. A 1 μmol/L increase in plasma fluoride was associated with a 10.36 mL/min/1.73 m2 lower estimated glomerular filtration rate (95% CI: -17.50, -3.22; p = 0.05), a 0.29 mg/dL higher serum uric acid concentration (95% CI: 0.09, 0.50; p = 0.05), and a 1.29 mg/dL lower blood urea nitrogen concentration (95%CI: -1.87, -0.70; p < 0.001). A 1 mg/L increase in water fluoride was associated with a 0.93 mg/dL lower blood urea nitrogen concentration (95% CI: -1.44, -0.42; p = 0.007). Fluoride exposure may contribute to complex changes in kidney and liver related parameters among U.S. adolescents. As the study is cross-sectional, reverse causality cannot be ruled out; therefore, altered kidney and/or liver function may impact bodily fluoride absorption and metabolic processes.


Beyond simple tests of value: measuring addiction as a heterogeneous disease of computation-specific valuation processes.
Addiction is considered to be a neurobiological disorder of learning and memory because addiction is capable of producing lasting changes in the brain. Recovering addicts chronically struggle with making poor decisions that ultimately lead to relapse, suggesting a view of addiction also as a neurobiological disorder of decision-making information processing. How the brain makes decisions depends on how decision-making processes access information stored as memories in the brain. Advancements in circuit-dissection tools and recent theories in neuroeconomics suggest that neurally dissociable valuation processes access distinct memories differently, and thus are uniquely susceptible as the brain changes during addiction. If addiction is to be considered a neurobiological disorder of memory, and thus decision-making, the heterogeneity with which information is both stored and processed must be taken into account in addiction studies. Addiction etiology can vary widely from person to person. We propose that addiction is not a single disease, nor simply a disorder of learning and memory, but rather a collection of symptoms of heterogeneous neurobiological diseases of distinct circuit-computation-specific decision-making processes.


Enhanced Heart Rate Prediction Model Using Damped Least-Squares Algorithm.
Monitoring a patient's vital signs is considered one of the most challenging problems in telehealth systems, especially when patients reside in remote locations. Companies now use IoT devices such as wearable devices to participate in telehealth systems. However, the steady adoption of wearables can result in a significant increase in the volume of data being collected and transmitted. As these devices run on limited battery power, they can run out of power quickly due to the high processing requirements of the device for data collection and transmission. Given the importance of medical data, it is imperative that all transmitted data adhere to strict integrity and availability requirements. Reducing the volume of healthcare data and the frequency of transmission can improve a device's battery life via an inference algorithm. Furthermore, this approach creates issues for improving transmission metrics related to accuracy and efficiency, which are traded-off against each other, with increasing accuracy reducing efficiency. This paper demonstrates that machine learning (ML) can be used to overcome the trade-off problem. The damped least-squares algorithm (DLSA) is used to enhance both metrics by taking fewer samples for transmission whilst maintaining accuracy. The algorithm is tested with a standard heart rate dataset to compare the metrics. The results showed that the DLSA provides the best performance, with an efficiency of 3.33 times for reduced sample data size and an accuracy of 95.6%, with similar accuracies observed in seven different sampling cases adopted for testing that demonstrate improved efficiency. This proposed method significantly improve both metrics using ML without sacrificing one metric over the other compared to existing methods with high efficiency.


Racial/Ethnic and Socioeconomic Disparities in Mental Health in Arizona.
Mental health issues are a rapidly increasing problem in the US. Little is known about mental health and healthcare among Arizona's Hispanic population. We assess differences in mental health service need, mental health diagnoses, and illicit drug use among 7,578 White and Hispanic participants in the 2010 Arizona Health Survey. Prevalence of mild, moderate, or severe psychological distress was negatively associated with SES among both Whites and Hispanics. Overall, Hispanics were less likely than Whites to have been diagnosed with a mental health condition; however, diagnosis rates were negatively associated with SES among both populations. Hispanics had considerably lower levels of lifetime illicit drug use than their White counterparts. Illicit drug use increased with SES among Hispanics but decreased with SES among Whites. After adjustment for relevant socio-demographic characteristics, multivariable linear regression suggested that Hispanics have significantly lower Kessler scores than Whites. These differences were largely explained by lower Kessler scores among non-English proficient Hispanics relative to English-speaking populations. Moreover, logistic regression suggests that Hispanics, the foreign born, and the non-English language proficient have lower odds of lifetime illicit drug use than Whites, the US born, and the English-language proficient, respectively. The unique social and political context in Arizona may have important but understudied effects on the physical and mental health of Hispanics. Our findings suggest mental health disparities between Arizona Whites and Hispanics, which should be addressed via culturally- and linguistically tailored mental health care. More observational and intervention research is necessary to better understand the relationship between race/ethnicity, socioeconomic status, healthcare, and mental health in Arizona.


AAV-mediated intravitreal gene therapy reduces lysosomal storage in the retinal pigmented epithelium and improves retinal function in adult MPS VII mice.
The beta-glucuronidase-deficient mucopolysaccharidosis type VII (MPS VII) mouse accumulates partially degraded glycosaminoglycans in many cell types, including retinal pigmented epithelial (RPE) cells in the eye. This lysosomal storage in RPE cells leads to progressive retinal degeneration and reduced function as measured by flash electroretinography (ERG). The impact of AAV-mediated intraocular gene therapy on pathology and retinal function was examined in normal and MPS VII mice treated at 4 weeks of age, when lysosomal storage is evident but functional impairment is minimal in affected animals. At 16 weeks, an age at which untreated MPS VII mice have advanced histologic lesions and significantly reduced ERG amplitudes, treated eyes had nearly normal levels of beta-glucuronidase activity, preservation of cells in the outer nuclear layer of the retina, and decreased lysosomal storage within the RPE. The AAV-treated MPS VII mice also had significantly increased dark-adapted ERG amplitudes compared to untreated MPS VII mice. Although retinal function was improved, the efficacy of the treatment depended heavily on parameters related to the injection procedure, such as the injection volume, injection site, and vector dose. These data suggest that intraocular AAV-mediated therapy may be efficacious for treating the retinal disease associated with certain lysosomal storage diseases.


Response of Nanodot Optically Stimulated Luminescence Dosimeters to Therapeutic Electron Beams.
Response of Al2O3:C-based nanoDot optically stimulated luminescence (OSL) dosimeter was studied for the dosimetry of 6, 9, 12, 16, and 20 MeV therapeutic electron beams. With reference to ionization chamber, no change in the response was observed with the change in the energy of electron beams for the field size from 6 cm × 6 cm to 25 cm × 25 cm, dose rates from 100 MU/min to 600 MU/min, and the linearity in the response up to 300 cGy. The fading of the transient signal was higher for 20 MeV electron beam than that of 6 MeV electron beam by about 5% as compared to value at 20 min after irradiation. The depletion of OSL signal per readout in 200 successive readouts was also found to change with dose and energy of electron beam from 6 MeV (9% and 12% per readout at 2 and 10 Gy, respectively) to 20 MeV (9% and 16% at 2 and 10 Gy, respectively). The OSL sensitivity changed in the range from 2% to 6% with accumulated doses from 2 to 8 Gy and with electron energy from 6 to 20 MeV, but the sensitivity could be reset using an optical annealing treatment. Although negligible fading for postirradiation storage from 20 min to several months, acceptable precision and linearity in the desired range, and high reproducibility makes nanoDot dosimeters very attractive for the dosimetry of therapeutic electron beams, a note should be made for changes in sensitivity at doses beyond 2 Gy and electron beams energy dependence in reuse, short-term fading, and signal depletion on repeated readout.


Efficacy of primary hepatitis B immunization in children with acute lymphoblastic leukemia.
Children with acute lymphoblastic leukemia (ALL) carry a high risk of hepatitis B virus (HIV) infection. The present study was conducted to see if prior routine hepatitis B vaccine received as a part of national immunization program could prevent HBV infection in these children. Ninety-six children with ALL were screened for HBV. Children were divided into three groups according to their initial HBV serology; previously vaccinated children (Group I) (n=34) previously unvaccinated and seronegative children (Group II) (n=56),and unvaccinated but HBsAg negative and anti-HBs positive children (group III) (n=6). Sixty-seven of 96 (69.7%) children received vaccination. The schedule was initiated during the third month of maintenance therapy and each course consisted of three doses given at one month interval. Anti-HBs seroconversion following the first course of three doses of hepatitis B vaccination in group I, II and III was 57%, 33% and 100%, respectively. It increased to 97% in Group I, 62.5% in Group II, 100% in Group III. HBsAg positivity was found in 11 children (11.5%) and all of them developed chronic hepatitis B. Ten of them were in Group II whereas only one child was in Group I (P<0.04). This data reveals that routine HBV vaccination within the national immunization program plays an important role in decreasing subsequent hepatitis B infection in children with ALL.


Novel easily available purine-based AIEgens with colour tunability and applications in lipid droplet imaging.
Recently, tetraphenylethene, triphenylamine and other man-made core AIE luminescent materials (AIEgens) have attracted significant scientific interest. However, the design and synthesis of natural product based, facile and color tunable AIEgens remains challenging. Herein, a novel series of AIEgens based on purine-core molecular rotors is reported, which can be facilely synthesized and shows color tunable emission. Moreover, these purine-based AIEgens exhibit lipid droplet specific properties in live cellular imaging with low background, high selectivity and excellent biocompatibility.


Revitalizing the typological approach: Some methods for finding types.
The purpose is to discuss and exemplify how a typological approach could be designed for studying phenomena believed to be best understood within a person-oriented theoretical framework. The focus is mainly restricted to the case of studying the typological structure of a sample at a single point in time, and restricted to analyzing variable profiles where each variable has a "negative" and "positive" endpoint. An artificial data set and an empirical data set were analyzed using two different methodological approaches, one more explorative (using LICUR, a cluster analysis-based procedure) and one more model-based (using the MCLUST procedure). For the artificial data set, the LICUR procedure was successful in finding the true classification structure but the MCLUST procedure performed surprisingly badly. For the empirical data set, both procedures produced rather similar solutions and they showed moderate validity. However, the LICUR solution appeared to be slightly superior. It was argued that applying a sound classification methodology and carefully validating the resulting classifications are extremely important, even more so in a developmental context. It was also argued that, in a number of situations, a more explorative approach could be more useful than a standard model-based one.


Placentome Nutrient Transporters and Mammalian Target of Rapamycin Signaling Proteins Are Altered by the Methionine Supply during Late Gestation in Dairy Cows and Are Associated with Newborn Birth Weight.
Background: To our knowledge, most research demonstrating a link between maternal nutrition and both fetal growth and offspring development after birth has been performed with nonruminants. Whether such relationships exist in large ruminants is largely unknown.Objective: We aimed to investigate whether increasing the methionine supply during late pregnancy would alter uteroplacental tissue nutrient transporters and mammalian target of rapamycin (mTOR) and their relation with newborn body weight.Methods: Multiparous Holstein cows were used in a randomized complete block design experiment. During the last 28 d of pregnancy, cows were fed a control diet or the control diet plus ethylcellulose rumen-protected methionine (0.9 g/kg dry matter intake) (Mepron; Evonik Nutrition & Care GmbH) to achieve a 2.8:1 ratio of lysine to methionine in the metabolizable protein reaching the small intestine. We collected placentome samples at parturition and used them to assess mRNA and protein expression and the phosphorylation status of mTOR pathway proteins.Results: Newborn body weight was greater in the methionine group than in the control group (44.1 kg and 41.8 kg, respectively; P ≤ 0.05). Increasing the methionine supply also resulted in greater feed intake (15.8 kg/d and 14.6 kg/d), plasma methionine (11.9 μM and 15.3 μM), and plasma insulin (1.16 μg/L and 0.81 μg/L) in cows during late pregnancy. As a result, mRNA expression of genes involved in neutral amino acid transport [solute carrier (SLC) family members SLC3A2, SLC7A5, SLC38A1, and SLC38A10], glucose transport [SLC2A1, SLC2A3, and SLC2A4], and the mTOR pathway [mechanistic target of rapamycin and ribosomal protein S6 kinase B1] were upregulated (P ≤ 0.07) in methionine-supplemented cows. Among 6 proteins in the mTOR pathway, increasing the methionine supply led to greater (P ≤ 0.09) protein expression of α serine-threonine kinase (AKT), phosphorylated (p)-AKT, p-eukaryotic elongation factor 2, and the p-mTOR:mTOR ratio.Conclusion: Supplemental methionine during late gestation increases feed intake and newborn body weight in dairy cows, and this effect may be mediated by alterations in the uteroplacental transport of nondispensable and dispensable amino acids and glucose at least in part through changes in gene transcription and mTOR signaling.


Antiviral innate immune memory in alveolar macrophages following SARS-CoV-2 infection.
Pathogen encounter results in long-lasting epigenetic imprinting that shapes diseases caused by heterologous pathogens. The breadth of this innate immune memory is of particular interest in the context of respiratory pathogens with increased pandemic potential and wide-ranging impact on global health. Here, we investigated epigenetic imprinting across cell lineages in a disease relevant murine model of SARS-CoV-2 recovery. Past SARS-CoV-2 infection resulted in increased chromatin accessibility of type I interferon (IFN-I) related transcription factors in airway-resident macrophages. Mechanistically, establishment of this innate immune memory required viral pattern recognition and canonical IFN-I signaling and augmented secondary antiviral responses. Past SARS-CoV-2 infection ameliorated disease caused by the heterologous respiratory pathogen influenza A virus. Insights into innate immune memory and how it affects subsequent infections with heterologous pathogens to influence disease pathology could facilitate the development of broadly effective therapeutic strategies.


The effect of vagal neural crest ablation on the chick embryo cloaca.
The cloaca, the caudal limit of the avian gastrointestinal tract, acts as a collecting chamber into which the gastrointestinal, urinary, and genital tracts discharge. It is intrinsically innervated by the enteric nervous system, which is derived from neural crest emigres that migrate from the vagal and sacral regions of the neural tube. Abnormal cloacal development can cause a number of anorectal anomalies, including persistent cloaca. Ablation of the vagal neural crest has previously been shown to result in an aganglionic hindgut to the extent of the colorectum. The aim of our study was to investigate the effect of vagal neural crest ablation on the cloaca, the limit of the hindgut in the developing chick embryo. Chick embryos were incubated until the 10-12 somite stage. The vagal neural tube corresponding to the level of somites 3-6 was then ablated, and eggs were incubated until harvested on embryonic day 11 (E11). Whole chick embryos were fixed, embedded in paraffin, and sectioned. Immunohistochemistry was then carried out using the HNK-1 monoclonal antibody to label neural crest cells, and results were assessed by light microscopy. Vagal neural crest ablation resulted in a dramatic decrease in the number of neural crest cells colonizing the chick embryo cloaca compared with control embryos. Ablated embryos contained only a small number of HNK-1-positive neural crest cells, which were scattered within the myenteric plexus in a disorganised pattern. Hypoganglionosis was also evident in other regions of the hindgut in ablated embryos. Ablation of the vagal neural crest results in a hypoganglionic cloaca in addition to hypoganglionosis of the hindgut. These results suggest that the cloaca is largely innervated by vagal neural crest emigres. Further studies involving quail-chick chimeras to investigate the exact contribution provided by both vagal and sacral neural crest cells to the cloaca should increase our understanding of the pathophysiology of conditions like persistent cloaca.


Limited specificity and large overlap of the functions of the mouse RAR gamma 1 and RAR gamma 2 isoforms.
The RAR gamma gene generates two major isoforms, RAR gamma 1 and RAR gamma 2, which originate from two distinct promoters. We report here the engineering of mice lacking RAR gamma 1, but in which RAR gamma 2 is normally expressed. The effect of this null mutation has been compared with those previously described for RAR gamma 2 and all RAR gamma isoforms (total RAR gamma gene inactivation), both in single mutants and in double mutants bearing additional null mutations in their RAR alpha, RAR beta or RXR alpha genes. RAR gamma 1 mutants, but not RAR gamma 2 mutants, displayed a subset of the abnormalities exhibited by total RAR gamma null mutants (growth deficiency, abnormal cricoid cartilage and occasional cervical vertebra defects), suggesting that RAR gamma 1 is the main isoform mediating the corresponding RAR gamma functions. Interestingly, cricoid cartilage defects were also found in a fraction of heterozygote animals for the RAR gamma 1, RAR gamma or RAR alpha mutations, indicating that wild type levels of RARs are required for the normal morphogenesis of this structure. Compound RAR alpha/RAR gamma 1 and RAR alpha/RAR gamma 2 double null mutants exhibited only a small fraction of the defects found in RAR alpha/RAR gamma double null mutants. Moreover, these defects were often partially penetrant, or corresponded to a less severe form. However, they occurred preferentially in certain compound mutants, demonstrating that given isoforms mediate specific functions of RAR gamma in the context of a RAR alpha null background. In a RXR alpha null background, both RAR gamma 1 and gamma 2 isoform mutations resulted in increased severity of the RXR alpha null ocular phenotype. Together, the present observations indicate that the functions of the two RAR gamma isoforms overlap to a large extent, but also that each of these isoforms exhibits a limited functional specificity. Furthermore, the occurrence of morphological defects in heterozygote mutants for a single RAR isoform provides a basis for explaining the strong conservation of these isoforms during vertebrate evolution.


Emergence of blaCTX-M-15, blaTEM-169 and blaPER-1 extended-spectrum β-lactamase genes among different Salmonella enterica serovars from human faecal samples.
Broad-spectrum β-lactams are used for empirical therapy of severe infections with non-typhoid Salmonella serotypes; however, activities of these drugs against the strains producing different β-lactamase is not so clear. This study investigated the prevalence of β-lactamase genes among isolates of S. enterica serovars from human faecal samples and determined their diversity in activity against different β-lactams. Antimicrobial resistance of faecal isolates of S. enterica to extended-spectrum cephalosporins was analysed and MIC values were determined for the strains presenting extended-spectrum β-lactamases (ESBLs) phenotypes. The β-lactamase genes were identified by PCR and sequencing. β-lactamase activity of the Salmonella strains exhibiting ESBL phenotype was detected by biological, iodometric, spectrophotometry and nitrocefin assays. Out of 202 S. enterica isolates, ESBLs phenotype was detected among 3.4% (7/202) of the strains. blaTEM-1 and blaCTX-M-15 were among the frequent β-lactamase genes. Detection of blaTEM-169 in S. enterica serovar Typhimurium and S. enterica serovar Bredeney and blaPER-1 in S. enterica serovar Infantis was a new finding in this experiment. Location of blaCTX-M-15/blaTEM-169/blaPER-1 genes on plasmid was confirmed in a transformation experiment. While crude extracts of the enzymes from each strain showed higher activity against cephalothin and cefotaxime, the lowest activity was detected against ceftazidime. The greatest synergistic activity was seen in a strain of S. enterica that carried blaCTX-M-15 and blaPER-1 genes compared with those presenting blaCTX-M-15/blaTEM-169 or blaCTX-M-15/blaTEM-1 genotypes. The results show dissemination of ESBLs encoding genes and their combined activity among different serovars of S. enterica that are a threat for future treatment options.


Changes in molecular order across the lamellar-to-inverted hexagonal phase transition depend on the position of the double-bond in mono-unsaturated phospholipid dispersions.
A series of mono-unsaturated phosphatidylethanolamine (PE) model membranes is studied using deuterium nuclear magnetic resonance (NMR) and differential scanning calorimetry. As the position of the double bond is systematically changed, the internal conformational motions are monitored through the bilayer-to-inverted-hexagonal phase transition. The order parameter profiles extracted from the NMR spectra report on the conformational order of the lipid and on the way this order is changed by structural reorganizations of the membrane. The calculation of a ratio of renormalized order parameter profiles is presented here as an attempt to distill the essential features of these changes into dimensionless descriptions of "shape" functions. This variation of the extent of molecular disorder along the long molecular axis of the phospholipids appears to be a recurring motif, modulated by temperature, structural rearrangement, and chemical composition of the membrane. The reported experimentally measured changes in the shape of the order parameter profile can be compared to those obtained during molecular dynamics simulation studies.


Social processes in communication and cognition in callitrichid monkeys: a review.
Studies of vocal development in nonhuman primates have found little evidence for plasticity in vocal production, somewhat more for usage of calls, with the greatest plasticity arising in response to calls of others. Generally, similar results were obtained with callitrichid monkeys, the marmosets and tamarins, but with several interesting exceptions. Infant pygmy marmosets show babbling behavior with improvement in adult call structure related to the amount and diversity of babbling. Adult marmosets alter call structure in response to changes in social partners, and wild marmosets have vocal dialects and modify call structure according to how far they are from other group members, suggesting the potential to modify call structure in different social and environmental contexts, though direct learning of novel vocalizations has not been observed. Infant cotton-top tamarins do not produce adult-like calls in appropriate contexts, at least in the first few months of life, but through food sharing from adults infants learn about appropriate foods and the appropriate contexts for food vocalizations. Tamarins modify call structure and usage with changes in social status. Tamarins, unlike other monkeys tested, can learn to avoid noxious foods through observation of other group members, and can learn about novel food locations. Recent studies provide evidence of contextual imitation in marmosets. The plasticity in vocal communication and evidence of social learning in marmosets and tamarins relative to other monkeys may be related to the cooperative breeding system of marmosets and tamarins. With a high degree of behavioral coordination among group members, there is a priority on monitoring signals and behavior of others and adjusting one's own signals and behavior. This creates the context for vocal plasticity and social learning.


Towards (re)conciliation: (re)constructing relationships between indigenous health workers and nurses.
Currently in Australia, both Aboriginal and non-Aboriginal Australians are attempting to reconcile themselves with a history of colonization/invasion, which resulted in human rights violations against Australia's indigenous people. Australian nursing has to examine its past in relation to the treatment of Aboriginal Australians. Relationships between nurses and Aboriginal health workers are the most commonly occurring professional relationships between nurses and Aboriginal people and are of key importance to the successful delivery of health services to Aboriginal communities. This qualitative study, grounded in feminism, aimed to explore the professional relationships between Aboriginal health workers and nurses and to develop insights which could assist the Australian nursing profession through a process of reconciliation with Aboriginal Australians. Feminist analysis of narrative text revealed several key themes as being crucial to this process. These were: learning to know and understand; towards workplace equity; and skill sharing -- learning from each other. Implications for nursing, in its journey toward reconciliation with Aboriginal Australians, are drawn from this study.


Bifidobacterium breve reduces apoptotic epithelial cell shedding in an exopolysaccharide and MyD88-dependent manner.
Certain members of the microbiota genus Bifidobacterium are known to positively influence host well-being. Importantly, reduced bifidobacterial levels are associated with inflammatory bowel disease (IBD) patients, who also have impaired epithelial barrier function, including elevated rates of apoptotic extrusion of small intestinal epithelial cells (IECs) from villi-a process termed 'cell shedding'. Using a mouse model of pathological cell shedding, we show that mice receiving Bifidobacterium breve UCC2003 exhibit significantly reduced rates of small IEC shedding. Bifidobacterial-induced protection appears to be mediated by a specific bifidobacterial surface exopolysaccharide and interactions with host MyD88 resulting in downregulation of intrinsic and extrinsic apoptotic responses to protect epithelial cells under highly inflammatory conditions. Our results reveal an important and previously undescribed role for B. breve, in positively modulating epithelial cell shedding outcomes via bacterial- and host-dependent factors, supporting the notion that manipulation of the microbiota affects intestinal disease outcomes.


Measurement of ionizing radiation using carbon nanotube field effect transistor.
Single-walled carbon nanotubes (SWNTs) are a new class of highly promising nanomaterials for future nano-electronics. Here, we present an initial investigation of the feasibility of using SWNT field effect transistors (SWNT-FETs) formed on silicon-oxide substrates and suspended FETs for radiation dosimetry applications. Electrical measurements and atomic force microscopy (AFM) revealed the intactness of SWNT-FET devices after exposure to over 1 Gy of 6 MV therapeutic x-rays. The sensitivity of SWNT-FET devices to x-ray irradiation is elucidated by real-time dose monitoring experiments and accumulated dose reading based on threshold voltage shift. SWNT-FET devices exhibit sensitivities to x-rays that are at least comparable to or orders of magnitude higher than commercial MOSFET (metal-oxide semiconductor field effect transistor) dosimeters and could find applications as miniature dosimeters for microbeam profiling and implantation.


Association of cystic fibrosis genetic modifiers with congenital bilateral absence of the vas deferens.
To investigate whether genetic modifiers of cystic fibrosis (CF) lung disease also predispose to congenital bilateral absence of the vas deferens (CBAVD) in association with cystic fibrosis transmembrane conductance regulator (CFTR) mutations. We tested the hypothesis that polymorphisms of transforming growth factor (TGF)-β1 (rs 1982073, rs 1800471) and endothelin receptor type A (EDNRA) (rs 5335, rs 1801708) are associated with the CBAVD phenotype. Genotyping of subjects with clinical CBAVD. Outpatient and hospital-based clinical evaluation. DNA samples from 80 subjects with CBAVD and 51 healthy male controls from various regions of Europe. This is one of the largest genetic studies of this disease to date. None. Genotype analysis. For single nucleotide polymorphism (SNP) rs 5335, we found increased frequency of the CC genotype among subjects with CBAVD. The difference was significant among Turkish patients versus controls (45.2% vs. 19.4%), and between all cases versus controls (36% vs. 15.7%). No associations between CBAVD penetrance and polymorphisms rs 1982073, rs 1800471, or rs 1801708 were observed. Our findings indicate that endothelin receptor type A polymorphism rs 5335 may be associated with CBAVD penetrance. To our knowledge, this is the first study to investigate genetic modifiers relevant to CBAVD.


The impact of personality factors on delay in seeking treatment of acute myocardial infarction.
Early hospital arrival and rapid intervention for acute myocardial infarction is essential for a successful outcome. Several studies have been unable to identify explanatory factors that slowed decision time. The present study examines whether personality, psychosocial factors, and coping strategies might explain differences in time delay from onset of symptoms of acute myocardial infarction to arrival at a hospital emergency room. Questionnaires on coping strategies, personality dimensions, and depression were completed by 323 patients ages 26 to 70 who had suffered an acute myocardial infarction. Tests measuring stress adaptation were completed by 180 of them. The patients were then categorised into three groups, based on time from onset of symptoms until arrival at hospital, and compared using logistic regression analysis and general linear models. No correlation could be established between personality factors (i.e., extraversion, neuroticism, openness, agreeableness, conscientiousness) or depressive symptoms and time between onset of symptoms and arrival at hospital. Nor was there any significant relationship between self-reported patient coping strategies and time delay. We found no significant relationship between personality factors, coping strategies, or depression and time delays in seeking hospital after an acute myocardial infraction.


Comparative effectiveness of multiple acupuncture therapies for primary insomnia: a systematic review and network meta-analysis of randomized trial.
Acupuncture has been widely practiced for primary insomnia (PI). However, the relative benefit and harm among acupuncture therapies remain uncertain. To compare and evaluate the effect differences of multiple acupuncture therapies for patients with PI. Systematic literature search for randomized controlled trials (RCTs). Pairs of reviewers independently conducted literature screening, data extraction, and risk of bias assessment. Meta-analysis was conducted using R and Stata software. The Grading of Recommendations Assessment, Development, and Evaluation system (GRADE) was used to assess certainty of evidence and interpret results. Fifty-seven RCTs with 4678 patients were included. Compared with usual treatment, multiple acupuncture therapies showed a better effect for Pittsburgh sleep quality index score. And acupoints catgut embedding (ACE) was proved to be the most effective with a moderate certainty of evidence; auricular acupressure or auricular acupuncture plus manual acupuncture (AP + MA), electroacupuncture plus acupoint application (EA + APA), and intradermal needle (IN) might be also the most effective with low certainty of evidence. ACE, ACE + MA, AP + MA, EA, EA + APA, HPN, MA and PBN + MA showed significantly improvement in effective rate when compared with usual treatment. Insufficient evidence reported on Epworth Sleepiness Scale, Athens Insomnia Scale, and recurrence rate. The most common slight adverse events mainly included hematoma, pain, headache, and bleeding. With moderate to low certainty of evidence, multiple acupuncture therapies showed impressive insomnia improvement, especially ACE, AP + MA, and EA + APA. Differences between therapies were small or insignificant and based-on low or very low certainty of evidence.


Genetic structure of the indigenous populations of Siberia.
This study explores the genetic structure of Siberian indigenous populations on the basis of standard blood group and protein markers and DNA variable number of tandem repeats (VNTR) variation. Four analytical methods were utilized in this study: Harpending and Jenkin's R-matrix; Harpending and Ward's method of correlating genetic heterozygosity (H) to the distance from the centroid of the gene frequency array (rii); spatial autocorrelation, and Mantel tests. Because of the underlying assumptions of the various methods, the numbers of populations used in the analyses varied from 15 to 62. Since spatial autocorrelation is based upon separate correlations between alleles, a larger number of standard blood markers and populations were used. Fewest Siberian populations have been sampled for VNTRs, thus, only a limited comparison was possible. The four analytical procedures employed in this study yielded complementary results suggestive of the effects of unique historical events, evolutionary forces, and geography on the distribution of alleles in Siberian indigenous populations. The principal components analysis of the R-matrix demonstrated the presence of populational clusters that reflect their phylogenetic relationship. Mantel comparisons of matrices indicate that an intimate relationship exists between geography, languages, and genetics of Siberian populations. Spatial autocorrelation patterns reflect the isolation-by-distance model of Malecot and the possible effects of long-distance migration.


Stages of intestinal carcinogenesis and their control by host NK cells. Implications to prevention and therapy.
DMH-induced (25mg/kg/wk) rodent intestinal carcinogenesis was re-examined using histometry under various conditions including modulation of host natural killer (NK) cell activity and mutagen administration. Early lesion was enterocyte hyperplasia all along the intestinal tract. Cell kinetic analysis showed that it was caused by a slightly altered but still functional "initiated" enterocyte population added to the normal population which remained unchanged. Primarily, initiated stem (IS) cells were added which then produced their initiated progeny which still renewed normally. NK cells countered the initiated cells selectively, either inhibiting IS cell proliferation or killing them when activated by lymphokines. Evidence was obtained that the IS cells could further transform into preneoplastic stem (PS) and then neoplastic stem (NS) cells under the influence of promoters and mutagens, respectively. Subsequent transformation of normal stem cells into IS, PS, and NS cells apparently is the basis of carcinogenesis. These produce lesions, hyperplastic, preneoplastic, and neoplastic, respectively, only when and where NK activity is inhibited. Under the influence of normal NK cells, they remain dormant (nonproliferative), compatible with normal life.


Apoptosis characterization in mononuclear blood leukocytes of HIV patients during dengue acute disease.
Dengue virus (DENV) co-circulation in Brazil represents a challenge for treatment and vaccine development. Despite public health impact, the occurrence of coinfections with other viruses is a common event. Increased T cell activation and altered inflammatory response are found during DENV coinfection with Human Immunodeficiency Virus (HIV) impacting HIV-pathogenesis. Even with Antiretroviral therapy (ART), HIV- treated patients had chronic immune activation and lymphocyte apoptosis. However, apoptotic mechanisms have not been investigated during coinfection with DENV. Our attention was attracted to apoptotic cell markers expressions in PBMCs from DENV and DENV/HIV coinfected patients. We found CD4/CD8 ratio inversion in most coinfected patients. CD4 T and CD8 T-cell subsets from DENV and DENV/HIV groups expressed low levels of anti-apoptotic protein Bcl-2. Furthermore, CD8 CD95 double positive cells frequency expressing low levels of Bcl-2 were significantly higher in these patients. Additionally, the density of Bcl-2 on classical monocytes (CD14++CD16-) was significantly lower during DENV infection. Upregulation of pro-apoptotic proteins and anti-apoptotic proteins were found in DENV and DENV/HIV, while catalase, an antioxidant protein, was upregulated mainly in DENV/HIV coinfection. These findings provide evidence of apoptosis triggering during DENV/HIV coinfection, which may contribute to knowledge of immunological response during DENV acute infection in HIV-patients treated with ART.


Progress in the treatment of childhood diabetes mellitus and obesity.
The 61st Annual Meeting and Scientific Sessions of the American Diabetes Association (ADA) in Philadelphia, PA, (June 22-26, 2001) presented many topics of interest to pediatric clinicians. Of particular interest were the results of the insulin injection arm of the Diabetes Prevention Trial for type 1 diabetes mellitus (DM) (DPT-1). Over 80,000 relatives of patients with type 1 DM were screened. Ultimately, 339 subjects were randomized either to active therapy (twice daily insulin injections plus an annual insulin infusion) or to close observation. Risk prediction algorithms appeared to be accurate. Unfortunately, however, insulin therapy did not decrease the risk of developing DM. Of note, this was primarily a pediatric study with most of those randomized under 21 years of age. As expected, young subjects (<12 years) progressed toward the development of DM at a faster rate than older subjects (>15 years). The second arm of the DPT-1 trial, testing oral insulin in those with intermediate risk (25-50%) for DM, is still recruiting subjects. The controversial topic of continuous subcutaneous insulin infusion (CSII) in young children was also addressed. Many investigators presented data strongly supporting the successful use of infusion pumps in young children. In general, glycemic control was improved or remained stable, the incidence of severe hypoglycemia was low, and families reported more flexibility in their lifestyle. Obesity, an increasing problem in pediatric patients, was also addressed.


On the mechanism of crystalline polymorph selection by polymer heteronuclei.
The phase-selective crystallization of acetaminophen (ACM) using insoluble polymers as heteronuclei was investigated in a combined experimental and computational effort to elucidate the mechanism of polymer-induced heteronucleation (PIHn). ACM heteronucleates from supersaturated aqueous solution in its most thermodynamically stable monoclinic form on poly(n-butyl methacrylate), whereas the metastable orthorhombic form is observed on poly(methyl methacrylate). When ACM crystals were grown through vapor deposition, only the monoclinic polymorph was observed on each polymer. Each crystallization condition leads to a unique powder X-ray diffraction pattern with the major preferred orientation corresponding to the crystallographic faces in which these crystal phases nucleate from surfaces of the polymers. The molecular recognition events leading to these outcomes are elucidated with the aid of computed polymer-crystal binding energies using docking simulations. This investigation illuminates the mechanism by which phase selection occurs during the crystallization of ACM using polymers as heteronuclei, paving the way for the improvement of methods for polymorph selection and discovery based on heterogeneous nucleation promoters.


Distillation-based Chinese Food Ingredient Recognition and Nutrition Estimation System.
The foods' ingredients and nutrition are of great significance for human health so that people can meet their fitness needs or avoid consuming allergenic and post-operative contraindicated foods. However, the diversity of recipes and the randomness of combinations in Chinese cuisine make great challenges for Chinese food identification. To address the above issues, we built a new lightweight end-to-end food query and nutrition recognition system, which is based on knowledge distillation and deep learning methods. Firstly, well-performed DenseNet-121 is used to recognize the categories of food. At the same time, ResNet-50 is used as the Net-T, and pre-trained VGG-16 is used as the Net-S in the knowledge distillation framework, which is used to recognize the ingredients of the food. Finally, ingredient nutrition is obtained by querying the ingredient table. Experiments illustrate the good performance of the proposed method, with 91.65% Accuracy of food classification and 92.01% Accuracy of ingredients recognition.


Impact of hepatitis B virus and hepatitis C virus infection on sperm parameters of infertile men.
Viral hepatitis is one of the health problems which have the effects on the health issues. It seems that hepatitis B virus (HBV) and hepatitis C virus (HCV) infection have negative impacts on the semen quality and male infertility rate. In this study, we aimed to evaluate the effects of HBV and HCV on sperm quality among Iranian infertile men referred to Royan Institute Reproductive Biomedicine Research Center between 2003 and 2014. This retrospective case-control study included 112 HBV positive infertile men and 47 HCV positive infertile men as case group and 112 HBV negative and HCV negative matched infertile men as a control group. All semen analysis and viral parameters assessment was performed in the central laboratory with the same method and instruments. Sperm count among infertile men with HBV and HCV infection was significantly lower than control group [the mean of the total sperm count 100.95░±░118.59, 118.22░±░141.18, 166.27░±░151.25 (p░<░0.001)]. Sperm motility was significantly decreased in HBV and HCV positive men in comparison to the control group [30.97░±░25.88, 31.09░±░28.72, 40.87░±░23.37, respectively (p░<░0.007)]. The percentage of normal sperm morphology was significantly higher in control group in comparison to HBV and HCV infected group [the mean of the normal semen morphology 3.23░±░3.27, 3.70░±░3.83, 4.51░±░3.15 p░<░0.015]. Although there is a significant decline in liquefaction time in the case group but the viscosity, semen volume, and PH of semen samples were similar in the both case and control groups. Our results suggest that HBV and HCV infection are associated with poor sperm quality.


Developmental regulation of intermediate filament and actin mRNAs during myogenesis is disrupted by oncogenic ras genes.
Terminal differentiation of skeletal myoblasts is accompanied by down-regulation of vimentin and beta-, gamma-actins and up-regulation of desmin and sarcomeric alpha-actin(s). To investigate whether the normal decline in expression of vimentin and beta-, gamma-actins was coupled to withdrawal of proliferating myoblasts from the cell cycle or was a direct consequence of terminal differentiation, expression of the mRNAs encoding the actin microfilaments and intermediate filaments was examined in differentiation-defective C2 myoblasts bearing oncogenic ras genes. When transferred to mitogen-deficient medium, myoblasts transfected with the valine-12 allele of the human Harvey (H)-ras gene ceased dividing but failed to appropriately down-regulate vimentin and beta-, gamma-actin mRNAs. On the contrary, the level of vimentin mRNA expression was increased about 2.5-fold. Conversely, alpha-sarcomeric actin and desmin mRNAs continued to be expressed at basal levels in ras-transfected myoblasts that had withdrawn from the cell cycle. The ability of the ras oncogene to interfere with developmental regulation of actin and intermediate filament mRNAs was dependent upon mutational activation and was not observed in myoblasts transfected with proto-oncogenic ras alleles. These results demonstrate that, in addition to preventing up-regulation of muscle-specific gene products during myogenesis, the oncogenic forms of ras interfere with the mechanism(s) responsible for down-regulation of genes whose expression declines during myoblast fusion.


Intoxication of epithelial cells by plasmid-encoded toxin requires clathrin-mediated endocytosis.
It has been shown that the autotransporter plasmid-encoded toxin (Pet) of enteroaggregative Escherichia coli (EAEC) produces cytotoxic and enterotoxic effects. Both effects can be explained by the proteolytic activity of Pet on its intracellular target alpha-fodrin (alphaII spectrin). In addition, Pet cytotoxicity and enterotoxicity depend on Pet serine protease activity, and on its internalization into epithelial cells. However, the mechanisms of Pet uptake by epithelial cells are unknown. Here, we show that Pet interacts with the plasma membrane of epithelial cells, and afterwards is detected inside the cells. Furthermore, Pet was internalized via clathrin-mediated endocytosis, since its internalization was inhibited by monodansylcadaverine and sucrose, but not by filipin or methyl-beta-cyclodextrin, which are drugs that interfere with protein entry via a clathrin-independent pathway. Additionally, Pet was immunoprecipitated by anti-clathrin antibodies, but not by anti-caveolin antibodies. Moreover, small interfering RNA (siRNA), designed to knock out clathrin gene expression in HEp-2 cells, prevented Pet internalization, and thereby the Pet-induced cytotoxic effect. However, the use of siRNA to knock out caveolin expression had no effect on Pet internalization, and the cytotoxic effect was clearly observed. Together, these data indicate that Pet secreted by EAEC binds to the cell surface via an unknown receptor, to be taken up by clathrin-mediated endocytosis, and exert its toxic effect in the cytoplasm.


Interactions between alcohol- and opioid-induced suppression of rat testicular steroidogenesis in vivo.
To examine interactions between alcohol and endogenous opioids in their suppressive effects on rat testicular function, the opioid antagonist naltrexone or the opioid agonist morphine was administered to adult male rats alone or in combination with alcohol. Serum testosterone, testicular interstitial fluid (TIF) testosterone, and TIF volumes were measured to assess testicular function. Naltrexone induced dose-dependent increases in serum and TIF testosterone levels without changes in TIF volume. Alcohol (0.5 g/kg) inhibited naltrexone-induced stimulation of testosterone secretion and shifted the naltrexone dose-response curve to the right. Conversely, naltrexone (0.05 mg/kg) inhibited alcohol-induced suppression of testosterone secretion and shifted the alcohol dose-response curve to the right. Relatively high doses of naltrexone (5 to 30 mg/kg) were needed to stimulate testosterone secretion maximally in rats treated with a low dose of alcohol (0.5 g/kg) and to stimulate normal levels of testosterone secretion in rats treated with a high dose of alcohol (2 g/kg). In addition, combined treatment with 1 and 30 mg/kg of naltrexone and 0.5 to 2 g/kg of alcohol did not alter blood alcohol concentrations significantly, suggesting that the interactions between alcohol and naltrexone were unrelated to gross changes in alcohol metabolism or bioavailability factors. Simultaneous treatments with a low dose of alcohol (0.3 g/kg), near the threshold of efficacy, and low-moderate doses of morphine (0.3 to 3 mg/kg) were not additive in suppressing testosterone secretion, compared with either agent alone. These results support the hypothesis that opioid antagonists can reverse the suppressive effect of alcohol on testicular steroidogenesis, but the results also suggest that endogenous opioids do not exclusively mediate alcohol's effects on testosterone secretion.


Laminin-3B11, a novel vascular-type laminin capable of inducing prominent lamellipodial protrusions in microvascular endothelial cells.
The basement membrane (BM) proteins laminins, which consist of α, β, and γ chains, support tissue structures and cellular functions. To date only α4 and α5 types of laminins have been identified in the BMs of blood vessels. Our recent study suggested the presence of novel α3B-containing laminins in vascular BMs. Here we identified and characterized the third member of vascular laminins, laminin-3B11 (Lm3B11). RT-PCR analysis showed that microvascular endothelial (MVE) cells and umbilical vein endothelial cells expressed the messages for the α3B, β1, β2, and γ1 chains. In the culture of MVE cells, α3B was associated with β1 and γ1, producing Lm3B11. Recombinant Lm3B11 was overexpressed by introducing the cDNAs of the three chains into HEK-293 cells and purified to homogeneity. Purified Lm3B11 exhibited relatively weak cell adhesion activity through both α3β1 and α6β1 integrins. Most characteristically, Lm3B11 strongly stimulated MVE cells to extend many lamellipodial protrusions. This pseudopodial branching was blocked by an inhibitor for Src or phosphatidylinositol 3-kinase. Consistently, Lm3B11 stimulated the phosphorylation of Src and Akt more strongly than other laminins, suggesting that the integrin-derived signaling is mediated by these factors. The unique activity of Lm3B11 appears to be favorable to the branching of capillaries and venules.


Variability of state school-based hearing screening protocols in the United States.
The prevalence of hearing loss has increased among United States children. As schools commonly perform hearing screens, we sought to contrast current United States school-based hearing screening protocols. State department of health and education websites were reviewed to assess school hearing screening protocols for the fifty states and the District of Columbia. Individuals listed on these websites were contacted as necessary to confirm and/or acquire relevant data. School-based hearing screening is currently required in 34/51 (67 %) states. Of these 34 states, 28 (82 %) mandate grades for screening, but only 20 (59 %) require screening beyond 6th grade. Pure tone audiometry is the most common screening method (33/34 [97 %]). A majority of states screen at 1, 2 and 4 kHz usually at 20 or 25 dB hearing level. Six states recommend or require testing at 6 or 8 kHz, which is necessary to detect high-frequency hearing loss. The results indicate that United States school-based hearing screens vary significantly. They focus on low frequencies with few testing adolescents for whom high-frequency hearing loss has increased. Disparities in hearing loss detection are likely, particularly considering the evolution of hazardous noise exposures and rising prevalence of hearing loss.


Suppression of preadipocyte differentiation and promotion of adipocyte death by HIV protease inhibitors.
Many human immunodeficiency virus (HIV)-infected patients taking combination antiretroviral therapy that includes HIV protease inhibitors experience atrophy of peripheral subcutaneous adipose tissue. We investigated the effects of HIV protease inhibitors on adipogenesis and adipocyte survival using the 3T3-L1 preadipocyte cell line. Several HIV protease inhibitors were found either to inhibit preadipocyte differentiation or to promote adipocyte cell death. One protease inhibitor, nelfinavir, elicited both of these effects strongly. When induced to differentiate in the presence of nelfinavir, 3T3-L1 preadipocytes failed to accumulate cytoplasmic triacylglycerol and failed to express normal levels of the adipogenic transcription factors CCAAT/enhancer-binding protein alpha and peroxisome proliferator-activated receptor gamma. The level of the proteolytically processed, active 68-kDa form of sterol regulatory element-binding protein-1, a transcription factor known to promote lipogenic gene expression, also was reduced markedly in nelfinavir-treated cells, whereas the level of the 125-kDa precursor form of this protein was unaffected. The inhibitory effect of nelfinavir occurred subsequent to critical early events in preadipocyte differentiation, expression of CCAAT/enhancer-binding protein beta and completion of the mitotic clonal expansion phase, because these events were unaffected by nelfinavir treatment. In addition, nelfinavir treatment of fully differentiated 3T3-L1 adipocytes resulted in DNA strand cleavage and severe loss of cell viability. In contrast, cell proliferation and viability of preadipocytes were unaffected by nelfinavir treatment. Thus, molecular or cellular changes that occur during acquisition of the adipocyte phenotype promote susceptibility to nelfinavir-induced cell death. When considered together, these results suggest that nelfinavir may promote adipose tissue atrophy by compromising adipocyte viability and preventing replacement of lost adipocytes by inhibiting preadipocyte differentiation.


Antidiabetic medication and risk of dementia in patients with type 2 diabetes: a nested case-control study.
Diabetes is a risk factor for dementia, but whether antidiabetic medication decreases the risk is unclear. We examined the association between antidiabetic medication and dementia. We performed a nested case-control study within a cohort of all 176 250 patients registered with type 2 diabetes in the Danish National Diabetes Register between 1995 and 2012. This population was followed for dementia diagnosis or anti-dementia medication use until May 2018. Using risk-set sampling, each dementia case (n = 11 619) was matched on follow-up time and calender year of dementia with four controls randomly selected among cohort members without dementia (n = 46 476). Ever use and mean daily defined dose of antidiabetic medication was categorized in types (insulin, metformin, sulfonylurea and glinides combined, glitazone, dipeptidyl peptidase 4 (DPP4) inhibitors, glucagon-like peptide 1 (GLP1) analogs, sodium-glucose transport protein 2 (SGLT2) inhibitors and acarbose). Conditional logistic regression models were fitted to calculate odds ratios (ORs) for dementia associated with antidiabetic medication use, adjusting for potential confounders. Use of metformin, DPP4 inhibitors, GLP1 analogs, and SGLT2 inhibitors were associated with lower odds of dementia after multible adjustments (ORs of 0.94 (95% confidence interval (CI): 0.89-0.99), 0.80 (95% CI 0.74-0.88), 0.58 (95% CI: 0.50-0.67), and 0.58 (95% CI: 0.42-0.81), respectively), with a gradual decrease in odds of dementia for each increase in daily defined dose. Analyses of the most frequent treatment regimes did not show any synergistic effects of combined treatment. Use of metformin, DPP4 inhibitors, GLP1 analogs and SGLT2 inhibitors was associated with lower risk of dementia in patients with diabetes.


Provision of enabling technology in professional sports.
Multiple-round golf tournaments are designed intentionally to separate individuals' scores as play proceeds. Variance analyses and consideration of individual differences (vs group mean effects) for a sample of professional events confirm that 3-, 4-, and 5-round tournaments show significantly increased variability (though stable means) from first to last rounds. It is argued here that the dispersion of scores increases as play proceeds because the more physically or mentally fit players emerge and continue to perform best. Furthermore, a marginal income analysis indicates that the average gain in earnings from a one-shot improvement in score is approximately $8,000. An interpretation based on fatigue, competition, and stress supports the Professional Golf Association's claim that provision of enabling devices, like a golf cart for disabled players, is also an enhancement and is thus unfair.


Primary structure and tissue distribution of the orphanin FQ precursor.
The heptadecapeptide orphanin FQ (OFQ) is a recently discovered neuropeptide that exhibits structural features reminiscent of the opioid peptides and that is an endogenous ligand to a G protein-coupled receptor sequentially related to the opioid receptors. We have cloned both the human and rat cDNAs encoding the OFQ precursor proteins, to investigate whether the sequence relationships existing between the opioid and OFQ systems are also found at the polypeptide precursor level, in particular whether the OFQ precursor would encode several bioactive peptides as do the opioid precursors, and to study the regional distribution of OFQ sites of synthesis. The entire precursor protein displays structural homology to the opioid peptide precursors, especially preprodynorphin and preproenkephalin. The predicted amino acid sequence of the OFQ precursor contains a putative signal peptide and one copy of the OFQ sequence flanked by pairs of basic amino acid residues. Carboxyl-terminal to the OFQ sequence, the human and rat precursors contain a stretch of 28 amino acids that is 100% conserved and thus may encode novel bioactive peptides. Two peptides derived from this stretch were synthesized but were found to be unable to activate the OFQ receptor, suggesting that if they are produced in vivo, these peptides would likely recognize receptors different from the OFQ receptor. To begin analyzing the sites of OFQ mRNA synthesis, Northern analysis of human and rat tissues were carried out and showed that the OFQ precursor mRNA is mainly expressed in the brain. In situ hybridization of rat brain slices demonstrated a regional distribution pattern of the OFQ precursor mRNA, which is distinct from that of the opioid peptide precursors. These data confirm that the OFQ system differs from the opioid system at the molecular level, although the OFQ and opioid precursors may have arisen from a common ancestral gene.