Dental Caries in 7-17-Year-Old Children in Moscow: A Clinical and a Questionnaire Study.
Sparse data is available concerning the distribution of decayed, extracted, filled/decayed, missing, filled tooth surfaces (defs/DMFS) and the impact of influencing risk factors in Moscow. We thus aimed to measure caries experience and to estimate its associations with relevant risk factors in schoolchildren. Data was obtained from 1004 schoolchildren aged 7-17. The clinical examination included the status of dental plaque, gingival bleeding and caries experience; defs/DMFS. The questionnaire was introduced to the children/parents, in order to measure socioeconomic and behavioural variables. The Fisher Exact test and chi-squared test were used to assess statistical significance of the distribution of the variables among groups. Bivariate and general estimating equations (GEE) analyses were applied to estimate the relative effect of the independent variables on the outcomes defined as median defs and median DMFS. The median defs and median DMFS varied among age groups. In the primary dentition, the bivariate analyses showed association between median defs and gender, plaque, toothache, self-satisfaction with the appearance of teeth, and intake of milk with sugar were associated (p <0.05). The multivariate analyses revealed that the median defs was lower in girls (OR = 0.9) and children with evidence of no plaque (OR = 0.7)/thin plaque (OR = 0.8), (p ≤ 0.002). In the permanent dentition, the bivariate analyses showed association between median DMFS and plaque, gingival bleeding, healthy dentition, use of toothpicks/dental floss, intake of biscuits etc, soft drinks and jam/honey, and education of the child's mother (p ≤ 0.02). Only gingival bleeding after probing (OR = 1.2) and higher education level of the mothers (OR = 0.9) were associated with the median DMFS in the multivariate analyses (p < 0.05). Clinical, socioeconomic and behavioural determinants were identified to influence caries in primary and permanent dentition in schoolchildren in Moscow. The findings might provide a reliable basis for improvements and education programmes in oral health promotion for children and adolescents.


Congenital Lobar Emphysema Presenting as Respiratory Distress in a Newborn.
Congenital lobar emphysema (CLE) is a rare developmental malformation that presents as neonatal respiratory distress and can be successfully managed with prompt intervention. Hyperinflation of the affected lobe with mediastinal shift is the characteristic radiological finding. However, the etiology mostly remains unknown. Here, we report a case of CLE that microscopically revealed bronchial cartilaginous hypoplasia as an underlying cause of this malformation.


The natural history of respiratory syncytial virus infection in cancer and transplant patients: implications for management.
Respiratory syncytial virus (RSV) has been reported to cause severe morbidity and mortality among cancer patients receiving chemotherapy with or without autologous peripheral blood stem cell transplantation (APBSCT). However, little is known about the natural history of this infection in these patients, and current standard practice, aerosolized ribavirin plus intravenous immunoglobulin (IVIG), is extremely expensive, difficult to use, and not supported by controlled clinical trials. The purpose of this observational study was to determine the frequency, seasonality, morbidity, and mortality of RSV infection in a group of cancer patients receiving cytotoxic chemotherapy with neither ribavirin nor IVIG treatment. During the period of October 3, 1997, through October 14, 1998, 190 cancer patients (median age, 58 years; 71 women) underwent viral nasopharyngeal washing prior to chemotherapy. Multiple myeloma (MM) accounted for most patients (147, 77%). RSV was recovered from cultures taken from 71 patients (37%) throughout the year, although more frequently during fall and winter seasons (P <.001) than spring and summer. Serious respiratory complications developed in 19 (27%) of 71 RSV-positive patients versus 24 (20%) of 119 patients whose RSV cultures were negative (P =.384). The presence of renal failure or increased lactate dehydrogenase (LDH) prior to chemotherapy and the development of mucositis were the only predictive factors for severe respiratory complications. Recovery of RSV from nasopharyngeal washings among cancer patients is common, occurs throughout the year, and does not appear to increase serious morbidity or mortality. RSV infection may not necessarily be a contraindication for APBSCT or an indication for therapy with aerosolized ribavirin and IVIG.


Vitamin K induces osteoblast differentiation through pregnane X receptor-mediated transcriptional control of the Msx2 gene.
Vitamin K is a fat-soluble vitamin that serves as a coenzyme for vitamin K-dependent carboxylase. Besides its canonical action, vitamin K binds to the steroid and xenobiotic receptor (SXR)/pregnane X receptor (PXR) and modulates gene transcription. To determine if the osteoprotective action of vitamin K is the result of the PXR/SXR pathway, we screened by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis the PXR/SXR target genes in an osteoblastic cell line (MC3T3-E1) treated with a vitamin K2 (menaquinone 4 [MK4]). Osteoblastic differentiation of MC3T3-E1 cells was induced by MK4. Msx2, an osteoblastogenic transcription factor, was identified as an MK4-induced gene. Functional analysis of the Msx2 gene promoter mapped a vitamin K-responsive element (PXR-responsive element [PXRE]) that was directly bound by a PXR/retinoid X receptor alpha heterodimer. In a chromatin immunoprecipitation analysis, PXR was recruited together with a coactivator, p300, to the PXRE in the Msx2 promoter. MK4-bound PXR cooperated with estrogen-bound estrogen receptor alpha to control transcription at the Msx2 promoter. Knockdown of either PXR or Msx2 attenuated the effect of MK4 on osteoblastic differentiation. Thus, the present study suggests that Msx2 is a target gene for PXR activated by vitamin K and suggests that the osteoprotective action of MK4 in the human mediates, at least in part, a genomic pathway of vitamin K signaling.


Effects of low confluency, serum starvation and hypoxia on the side population of cancer cell lines.
The cancer stem cell theory describes a small subset of cancer cells that have the ability to initiate and drive the growth of a tumor. The niche refers to the environmental factors and the surrounding cells within which the tumor develops. The exact relationship between cancer stem cells and the tumor niche is not known. However, using side population analysis by flow cytometry, it is possible to analyze the relationship between environmental stresses and putative cancer stem cells. The side population is a subpopulation of cells that efflux Hoechst 33342 and has been previously shown to be enriched for cancer stem cells. Using this technique, we characterized the response of side population cells to low confluency, serum starvation and hypoxia using three different human cancer cell lines. We found that these stresses, characteristic of the tumor niche enrich the side population of DLD1, SW480 and MCF7 cancer cell lines, thus possibly predisposing the tumor to a more malignant phenotype.


Self-regulation resources and physical activity participation among adults with type 2 diabetes.
Physical activity plays a crucial role in the prevention and treatment of type 2 diabetes. Therefore, it is important to understand why so few adults with type 2 diabetes regularly engage in physical activity. The role of self-regulation in the context of health-related behavior adherence, especially in terms of physical activity engagement and adherence, has largely been reviewed based on the strength energy model. Building on this line of research, the aim of this theoretical work was to highlight how self-regulation and ego depletion can influence the lower rate of physical activity participation among adults with type 2 diabetes, compared to adults from the general population.


Formation of a cleavasome: enhancer DNA-2 stabilizes an active conformation of NaeI dimer.
Cleavage of DNA by NaeI-type restriction enzymes is stimulated by a DNA element with affinity for the activator site of the enzyme: a cleavage-enhancer DNA element. Measurements of the mobility of NaeI activity in comparison with protein standards on gel permeation columns and glycerol gradients demonstrated that NaeI, without enhancer, can form a 70,000 MW dimer. The dimer, however, is inactive: it could not cleave the "resistant" NaeI site in M13mp18 DNA in the absence of enhancer. In cleavage assays, enhancer stimulated either DNA nicking or DNA cleavage, depending upon NaeI concentration, and reduced the NaeI concentration required for the transition from nicking to cleavage activity. A gel mobility-shift assay of the interaction of NaeI with enhancer showed the formation of two complexes. Results using different sized DNAs and different percentage acrylamide gels for gel mobility-shift analysis implied that the two complexes were caused by NaeI monomer and dimer structures rather than one and two DNA binding. Dimer formation increased with the affinity of enhancer for NaeI. UV cross-linking "captured" the NaeI-enhancer complex; electrophoretic analysis of the cross-linked products showed NaeI dimer bound to enhancer. These results imply a model for cleavage enhancement in which enhancer binding stabilizes an active NaeI dimer conformation ("cleavasome") that cleaves both DNA strands before dissociating.


Psychoanalysis and the Challenges of Disciplinary Exportation: Notes from the African "Periphery" on the Method/Theory Distinction.
This paper briefly reports on the current expansion of opportunities for clinical education in psychoanalysis in southern Africa, various regions of Asia, and Iran. It is a preliminary reconsideration of whether the disciplinary export of psychoanalysis is another egregious exercise in neo-colonialism, as contrasted with its possibly liberatory significance. The author argues that much of the discussion around the universality of many theoretical propositions needs to be reformulated. For example, the controversies over the Oedipus complex have typically been articulated in terms of the effects of specific social arrangements, familial structures, and styles of maternal and paternal functioning, rather than in terms of the universal-but with much cultural variation in its implementation-incest taboo. It is argued that dissemination of the psychoanalytic method (and the four coordinates that follow from it) may be desirable, but in ways the export of the hermeneutic assumptions of certain theoretical models is not.


Differential scanning fluorimetry measurement of protein stability changes upon binding to glycosaminoglycans: a screening test for binding specificity.
The interaction between glycosaminoglycans (GAGs) and proteins is important for the regulation of protein transport and activity. Here we present a novel method for the measurement of protein-GAG interactions suitable for high-throughput screening, able to discriminate between the interactions of a protein with GAGs of different structures. Binding of proteins to the GAG heparin, a proxy for sulfated regions of extracellular heparan sulfate, was found to enhance the stability of three test proteins, fibroblast growth factors (FGFs)-1, -2, and -18. Chemically modified heparins and heparin oligosaccharides of different lengths stabilized the three FGFs to different extents, depending on the pattern of sugar binding specificity. The method is based on a differential scanning fluorescence approach. It uses a Sypro Orange dye, which binds to exposed core residues of a denatured protein and results in an increased fluorescence signal. It is convenient, requiring low micromolar amounts of protein and ligand compared to other interaction assays, employing only a real-time polymerase chain reaction (PCR) instrument.


Network representation of protein interactions-Experimental results.
A graph theoretical analysis of nuclear magnetic resonance (NMR) data of six different protein interactions has been presented. The representation of the protein interaction data as a graph or network reveals that all of the studied interactions are based on a common functional concept. They all involve a single densely packed hub of functionally correlated residues that mediate the ligand binding events. This is found independent of the kind of protein (folded or unfolded) or ligand (protein, polymer or small molecule). Furthermore, the power of the graph analysis is demonstrated at the examples of the Calmodulin (CaM)/Calcium and the Cold Shock Protein A (CspA)/RNA interaction. The presented approach enables the precise determination of multiple binding sites for the respective ligand molecules.


Molecular characterization and clonal relationships among Escherichia coli strains isolated from broiler chickens with colisepticemia.
This study characterized 52 Escherichia coli isolates from distinct diseased organs of 29 broiler chickens with clinical symptoms of colibacillosis in the Southern Brazilian state of Rio Grande do Sul. Thirty-eight isolates were highly virulent and 14 were virtually avirulent in 1-day-old chicks, yet all isolates harbored virulence factors characteristic of avian pathogenic E. coli (APEC), including those related to adhesion, iron acquisition, and serum resistance. E. coli reference collection phylogenetic typing showed that isolates belonged mostly to group D (39%), followed by group A (29%), group B1 (17%), and group B2 (15%). Phylogenetic analyses using the Amplified Ribosomal DNA Restriction Analysis and pulse-field gel electrophoresis methods were used to discriminate among isolates displaying the same serotype, revealing that five birds were infected with two distinct APEC strains. Among the 52 avian isolates, 2 were members of the pandemic E. coli O25:H4-B2-ST131 clone.


Multiple Injuries to the Lower Urinary Tract: Two Cases and Comparison with the EAU Guidelines.
Blunt trauma to the lower urinary tract is usually associated with pelvic fractures. The European Association of Urology (EAU) provides guidelines to diagnose and treat these injuries. The guidelines summarise the available evidence and provide recommendations on diagnosis and treatment of these patients. Therefore, these guidelines are important adjuncts to the urologist and emergency physician in the clinical decision-making. However, strict adherence to the guidelines is not always easy or possible because of concomitant injuries obscuring the clinical picture. This is illustrated by two case reports of concomitant injuries of the lower urinary tract (bladder with urethral injury). The clinical decisions will be discussed point by point and should serve as a practical teaching moment for the reader.


[Limbal transplantation: multicenter retrospective case series analysis].
To report the results of limbal transplantation (LT) in patients with limbal stem cell deficiency (LSCD) in the context of ocular surface diseases. A multicenter (5 centers) retrospective case series analysis of patients who underwent LT between 1996 and 2004 was performed. Data were collected by the same researcher using a customized database. Success was defined by the absence of a persistent corneal epithelial defect, on-going inflammation or recurrence of a pterygium. Data from 72 LT performed in 61 patients (65 eyes) with a mean follow-up of 20.8 months (SD 23.5; range, 3-115) were analyzed. There were 33 males and 28 females with a mean age of 55.8 years (SD: 15.6; range, 20-89). Fifty-eight (80.6%) LT were autografts (40 pterygia, 12 alkali burns, 3 iatrogenic cases, 2 viral infections, 1 neoplasia case) and 14 (19.4%) were allografts from cadaveric donors (7 immune-based disorders, 6 alkali burns, 1 iatrogenic case); all patients receiving allografts also received systemic immunosuppression. Of the total number of LT, 48 (66.7%) were successful. This proportion increased to 81.0% (47/58) when autografts were used. However, only 7.1% (1/14) of all allografts were successful. The success rate was higher (80.0%) when performed for a pterygium and lower when done for immune-based inflammation (14.3%). Autograft tissue for LT is always preferable to allografts to surgically treat LSCD, as clinical success is significantly higher, and systemic immunosuppression is avoided. As expected, immune-based disorders are the most difficult cases to treat. LT has been shown to be an excellent option for recurrent pterygium, although prospective studies need to be performed to further corroborate these results.


Effect of Helicobacter pylori eradication on subsequent dysplasia development after endoscopic resection of gastric dysplasia.
Eradication of Helicobacter pylori reduces the incidence of gastric cancer, and may inhibit gastric dysplasia progression into gastric cancer. The aim of this study was to investigate the effect of eradication of Helicobacter on the incidence of subsequent gastric dysplasia development after endoscopic resection. Medical records of patients who underwent endoscopic resection for gastric dysplasia were retrospectively reviewed. Presence of H. pylori was assessed by the Campylobacter-like organism test and histology. The rate of subsequent dysplasia development after endoscopic resection between the eradication group and non-eradication group was compared. Total of 129 patients positive for H. pylori infection were included for analysis. Of these, 85 patients received successful eradication therapy and 44 patients did not receive eradication therapy or failed to achieve successful eradication. Sex, mean age and pathologic grade of dysplasia did not differ between the two groups. In univariate analysis, the grade of intestinal metaplasia (p=0.013) significantly differed between metachronous dysplasia group and non-metachrounous dysplasia group. In multivariate analysis, eradication of H. pylori (p=0.014) was related to reduced incidence of subsequent gastric dysplasia development after endoscopic resection. Eradication of H. pylori likely has a beneficial effect in preventing the development of subsequent gastric dysplasia, a premalignant lesion of gastric cancer, after endoscopic resection.


Connexin 43 Phosphorylation: Implications in Multiple Diseases.
Connexin 43 (Cx43) is most widely distributed in mammals, especially in the cardiovascular and nervous systems. Its phosphorylation state has been found to be regulated by the action of more than ten kinases and phosphatases, including mitogen-activated protein kinase/extracellular signaling and regulating kinase signaling. In addition, the phosphorylation status of different phosphorylation sites affects its own synthesis and assembly and the function of the gap junctions (GJs) to varying degrees. The phosphorylation of Cx43 can affect the permeability, electrical conductivity, and gating properties of GJs, thereby having various effects on intercellular communication and affecting physiological or pathological processes in vitro and in vivo. Therefore, clarifying the relationship between Cx43 phosphorylation and specific disease processes will help us better understand the disease. Based on the above clinical and preclinical findings, we present in this review the functional significance of Cx43 phosphorylation in multiple diseases and discuss the potential of Cx43 as a drug target in Cx43-related disease pathophysiology, with an emphasis on the importance of connexin 43 as an emerging therapeutic target in cardiac and neuroprotection.


Comparison of two public sector tertiary care hospitals' management in reducing direct medical cost burden on breast carcinoma patients in Lahore, Pakistan.
Breast cancer is one of the major causes of death incurring highest morbidity and mortality amongst women of Pakistan. The purpose of this study was to assess and compare the role of two public sector tertiary care hospitals' management in reducing out of pocket (OOP) expenses on direct medical costs borne by breast carcinoma patients' household from diagnosis through treatment. Moreover, the study intended to explore the reasons of opting private diagnostic facilities by the said patients during the services taken from the foresaid tertiary care centers. A purposive sample of 164 primary breast carcinoma patients was recruited for data collection of this cross-sectional study. Face to face interviews and semistructured questionnaires were adopted as method of data gathering tools. Major cost components of direct medical costs were used to compare the financial strain on the patients' households of both targeted hospitals. In addition, information was collected regarding the reasons of opting private diagnostic centers for investigations. Frequency, percentages, median and inter quartile range (IQR) were calculated for the data. Non-parametric variables were compared using the Mann-Whitney U test. It was observed that overall direct medical cost borne by the breast carcinoma patients' households in Jinnah hospital (median US$1153.93 / Rs. 118,589) was significantly higher than Mayo hospital (median US$427.93 /Rs. 43,978), p<0.001; r=0.623. Moreover, spending on almost all of the components of direct medical cost were found smaller in case of Mayo hospital's patients as compared to Jinnah hospital. This study indicates that OOP direct medical cost burden was found considerably less in Mayo hospital as compared to Jinnah hospital. The OOP expenditures on chemotherapy were overwhelmingly high. However, high spending on privately opted investigations procedures was the common issue of the patients under treatment in both hospitals.


Isomeric identification by laser control mass spectrometry.
The influence shaped femtosecond laser pulses have on molecular photofragmentation and ionization, coupled with the intrinsic sensitivity of mass spectrometry, results in a powerful tool for fast, accurate, reproducible and quantitative isomeric identification. Complex phase functions are introduced to enhance differences during the laser-molecule interactions, which depend on geometric structure, resulting in different fragmentation fingerprints. A full account is given on the setup and results leading to a technique that can be used to distinguish between compounds normally indistinguishable by conventional electron ionization mass spectrometry. We demonstrate geometric and structural isomer identification of cis-/trans-3-heptene, cis-/trans-4-methyl-2-pentene, o-/p-cresol and o-/p-xylene. For the positional isomers of xylene we present a complete dataset consisting of 1024 different phases to explore phase complexity. A selection of two phases from that data can then be used to achieve quantitative identification in mixtures of xylene isomers. Finally, we evaluate receiver operational curves obtained from our experimental data to demonstrate the reliability that can be achieved by femtosecond laser control mass spectrometry.


Glutaric aciduria type I: a neuroimaging diagnosis?
Glutaric aciduria type I is an autosomal recessive disorder of organic acid metabolism secondary to glutaryl-coenzyme A (CoA) dehydrogenase deficiency. We report a previously healthy 17-month-old girl who presented with acute dystonia. Conventional T2-weighted and fluid-attenuated inversion recovery magnetic resonance images of the brain showed hyperintensity in the caudates and putamina bilaterally with subtle involvement of the medial frontal lobes. Diffusion-weighted magnetic resonance images showed striking restricted diffusion in the caudates and putamina consistent with acute necrosis. Single-voxel hydrogen magnetic resonance spectroscopy of the involved areas was normal. The clinical diagnosis of glutaric aciduria type I was confirmed by elevation of 3-hydroxyglutaric and glutaric acids. Diffusion-weighted magnetic resonance imaging is a sensitive indicator of basal ganglia necrosis in glutaric aciduria type I.


FCGR2A as one novel potential target for poor survival prognosis of clear cell renal cell carcinoma.
Clear cell renal cell carcinoma (ccRCC) is the most common type of renal cell carcinoma. Immunoglobulin FcγRIIa receptor (FCGR2A) has been implicated in various cancers, however, its role on ccRCC is not well studied. A total of 151 patients with ccRCC were recruited for the study. Cox proportional hazards regression analysis was performed to calculate the hazard radios of FCGR2A expression and tumor characteristics. Pathological changes associated with ccRCC in tumor tissue sections were analyzed by hematoxylin-eosin staining. Immunohistochemical and immunofluorescence staining were used to detect the protein expression of FCGR2A in the tissue sections. Correlation between the expression of FCGR2A and the overall survival (OS) of ccRCC patients was analyzed by biological process neural network and support vector machine. The expression of FCGR2A was significantly correlated with the TNM of tumor, family history of ccRCC and Fuhrman stage of ccRCC. Patients with high FCGR2A expression in the tumor tissue, had poorer OS than the patients with low and moderate FCGR2A expression. The Receiver operating characteristic curve showed that FCGR2A can be used as a sensitive and specific biomarker for the diagnosis of ccRCC. Western blotting revealed that the FCGR2A was expressed at higher levels in the ccRCC tissues. Biological process neural network and support vector machine fitting showed that the R2 between FCGR2A and survival time of ccRCC patients was 0.8429 and 0.7669, respectively. FCGR2A is highly expressed in ccRCC, higher expression of FCGR2A is associated with poorer OS of ccRCC.


House dust mite sublingual immunotherapy: a double-blind, placebo-controlled study in elderly patients with allergic rhinitis.
Immunotherapy in elderly patients is controversial, and there is still no evidence supporting this treatment's safety and efficacy in this population. This study was performed to evaluate the safety and efficacy of specific sublingual immunotherapy for house dust mite (HDM) allergens in patients over 60 years of age with allergic rhinitis and a confirmed allergy to HDM. This study sought to assess nasal symptoms during HDM season, reduce medication use and monitor for adverse reactions during immunotherapy. One hundred and eleven 60- to 75-year-old patients with allergic rhinitis and a confirmed Dermatophagoides pteronyssinus and Dermatophagoides farinae allergy were included in the study. The patients were individually randomized to active or placebo groups using a double-blind method (NCTO01605760 ClinicalTrials.gov). A total of 51 subjects in the sublingual allergen-specific immunotherapy (SLIT) group (Staloral 300R, Stallergenes, France) and 57 subjects in the placebo group were monitored for 3 years. Forty-seven patients completed 3 years of SLIT, and 48 subjects finished the placebo treatment in the same time period. The total nasal symptom score decreased by 44% in the active group and 6% in the placebo group after 3 years of SLIT. This difference was only significant in the active group (P < 0.05). At the end of therapy, the total medication score of the active group decreased significantly by a maximum of 51% (P < 0.05), whereas the total medication score of the placebo control group showed an insignificant decrease (P = 0.56). There were no systemic adverse reactions during the study. Sublingual allergen-specific immunotherapy in elderly patients with a HDM allergy to D. pteronyssinus and D. farinae generated a significant clinical improvement in the active group compared with the placebo group, particularly during the heating season. This therapy was well tolerated.


Timing of Pubertal Development in Boys and Girls With Congenital Heart Defects: A Nationwide Cohort Study.
Background Children with congenital heart defects (CHD) have an increased risk of developmental delay. It remains sparsely investigated if these patients also have a delayed pubertal development. In this nationwide cohort study, we evaluated if CHD was associated with timing of puberty using longitudinally collected data on pubertal milestones. Methods and Results We used data from the Danish nationwide Puberty Cohort. Information on CHD was obtained from the Danish National Patient Register. Information on pubertal development was obtained from 15 780 children through questionnaires answered half-yearly from 11 years until 18 years or full maturity. Using a multivariable regression model for censored time-to-event data, mean difference in age at attaining each pubertal milestone was estimated, including a combined pubertal marker. Compared with children without CHD, analyses were performed for both CHD overall and subdivided into simple and complex CHD. In a subanalysis, analyses were repeated in children born at term. In total, 137 children (62 boys and 75 girls) had a CHD diagnosis. Overall, no difference in age at pubertal timing was observed for children with CHD compared with unaffected children. The average differences were small for both boys (1.6 [95% CI, -2.6 to 5.7] months) and girls (1.0 [95% CI, -2.5 to 4.4] months). The same differences were observed when subdividing into simple or complex CHD and when restricting to children born at term. Conclusions We found no association between CHD and pubertal timing. For the group of children with complex CHD, we were unable to exclude a later pubertal timing.


Three hundred and thirty cycles of preimplantation genetic diagnosis for serious genetic disease: clinical considerations affecting outcome.
To report on our experience with preimplantation genetic diagnosis (PGD) cycles performed for serious genetic disease in relation to the clinical factors affecting outcome. Retrospective review of data from a single centre. Tertiary referral PGD centre in a London teaching hospital. The PGD cycles included 172 cycles for chromosome rearrangements, 96 cycles for single-gene disorders and 62 cycles for X-linked disorders. In vitro fertilisation was the preferred method in chromosome rearrangement and X-linked cases, while intra cytoplasmic sperm injection was used in all single-gene disorders. Appropriate in situ hybridisation fluorescence probes were used in chromosome rearrangement and X-linked cases and polymerase chain reaction was used in single-gene disorders. All pregnancies were followed till delivery. Live birth rate per PGD cycle started. Eighty-six percent of cycles started (283) reached oocyte retrieval and 3743 eggs were collected, of which 2086 fertilised normally (55.7%). Two hundred and fifty cycles (76%) had embryos sutiable for biopsy on day 3 of in vitro culture, 1714 embryos were biopsied, and in 205 cycles (62%), there was at least one unaffected embryo available for transfer, resulting in 90 pregnancies, 68 clinical pregnancies and 58 live birth. The live birth rate was 18% per cycle started, 21% per egg retrieval and 28% per embryo transfer which significantly affected the live birth outcome. Woman age, number of eggs collected and achieving cryopreservation of surplus embryos had no statistically significant effect on treatment outcome. The live birth outcome of PGD cycles for serious genetic disorder is modest and is affected by the number of embryos genetically suitable for transfer.


The alcohol-related psychosocial and behavioral risks of a nationally representative sample of adolescents.
This study, a secondary analysis of the National Longitudinal Study of Adolescent Health, used a representative sample of 7th-through 12th-grade students enrolled in US public schools between April and December 1995. Data were collected in respondents' homes using trained interviewers. A subset of 4,485 adolescents aged 12-17 were surveyed with regard to alcohol-use practices and related health-risk behavior, interpersonal problems, and demographic characteristics. Results showed adolescent males as significantly more likely to drink at high risk than adolescent females. Among those who drank one or more times in the past year, older adolescents were significantly more likely to report high-risk drinking than younger adolescents. Significantly more high-risk adolescents reported having a hangover, vomiting, regretting a behavior, having trouble with parents, regretting a sexual activity, having dating problems, fighting, having trouble with friends, and experiencing school trouble than did low-risk adolescents. These findings underscore the long-range significance of a coordinated school health program; in particular, school health services, school health instruction, and school health environment. Implications for school-based and community-based prevention and intervention programs are presented.


Insights into ultra-low affinity lipase-antibody noncovalent complex binding mechanisms.
Detection of host cell protein (HCP) impurities is critical to ensuring that recombinant drug products, including monoclonal antibodies (mAbs), are safe. Mechanistic characterization as to how HCPs persist in drug products is important to refining downstream processing. It has been hypothesized that weak lipase-mAb interactions enable HCP lipases to evade drug purification processes. Here, we apply state-of-the-art methods to establish lipase-mAb binding mechanisms. First, the mass spectrometry (MS) approach of fast photochemical oxidation of proteins was used to elucidate putative binding regions. The CH1 domain was identified as a conserved interaction site for IgG1 and IgG4 mAbs against the HCPs phospholipase B-like protein (PLBL2) and lysosomal phospholipase A2 (LPLA2). Rationally designed mutations in the CH1 domain of the IgG4 mAb caused a 3- to 70-fold KD reduction against PLBL2 by surface plasmon resonance (SPR). LPLA2-IgG4 mutant complexes, undetected by SPR and studied using native MS collisional dissociation experiments, also showed significant complex disruption, from 16% to 100%. Native MS and ion mobility (IM) determined complex stoichiometries for four lipase-IgG4 complexes and directly interrogated the enrichment of specific lipase glycoforms. Confirmed with time-course and exoglycosidase experiments, deglycosylated lipases prevented binding, and low-molecular-weight glycoforms promoted binding, to mAbs. This work demonstrates the value of integrated biophysical approaches to characterize micromolar affinity complexes. It is the first in-depth structural report of lipase-mAb binding, finding roles for the CH1 domain and lipase glycosylation in mediating binding. The structural insights gained offer new approaches for the bioengineering of cells or mAbs to reduce HCP impurity levels.Abbreviations: CAN, Acetonitrile; AMAC, Ammonium acetate; BFGS, Broyden-Fletcher-Goldfarb-Shanno; CHO, Chinese Hamster Ovary; KD, Dissociation constant; DTT, Dithiothreitol; ELISA, Enzyme-linked immunosorbent assay; FPOP, Fast photochemical oxidation of proteins; FA, Formic acid; F(ab'), Fragment antibodies; HCP, Host cell protein; IgG, Immunoglobulin; IM, Ion mobility; LOD, Lower limit of detection; LPLA2, Lysosomal phospholipase A2; Man, Mannose; MS, Mass spectrometry; MeOH, Methanol; MST, Microscale thermophoresis; mAbs, Monoclonal antibodies; PPT1, Palmitoyl protein thioesterase; ppm, Parts per million; PLBL2, Phospholipase B-like protein; PLD3, Phospholipase D3; PS-20, Polysorbate-20; SP, Sphingomyelin phosphodiesterase; SPR, Surface plasmon resonance; TFA, Trifluoroacetic acid.


Photobiomodulation provides neuroprotection through regulating mitochondrial fission imbalance in the subacute phase of spinal cord injury.
Increasing evidence indicates that mitochondrial fission imbalance plays an important role in delayed neuronal cell death. Our previous study found that photobiomodulation improved the motor function of rats with spinal cord injury. However, the precise mechanism remains unclear. To investigate the effect of photobiomodulation on mitochondrial fission imbalance after spinal cord injury, in this study, we treated rat models of spinal cord injury with 60-minute photobiomodulation (810 nm, 150 mW) every day for 14 consecutive days. Transmission electron microscopy results confirmed the swollen and fragmented alterations of mitochondrial morphology in neurons in acute (1 day) and subacute (7 and 14 days) phases. Photobiomodulation alleviated mitochondrial fission imbalance in spinal cord tissue in the subacute phase, reduced neuronal cell death, and improved rat posterior limb motor function in a time-dependent manner. These findings suggest that photobiomodulation targets neuronal mitochondria, alleviates mitochondrial fission imbalance-induced neuronal apoptosis, and thereby promotes the motor function recovery of rats with spinal cord injury.


Admission of hypertensive patients at the University of Benin Teaching Hospital, Nigeria.
There is a suggestion of an identifiable impact of hypertension on all cause mortality in rural Africa. There is however paucity of hard data on the impact of morbidity or modality from this disease. To determine the contribution of hypertension (HT) to adult morbidity and mortality at the University of Benin Teaching Hospital (UBTH) in Benin City, Nigeria. Retrospective study. University of Benin Teaching Hospital. Adult hypertensive admissions (HTA) in Benin City, Edo state, Nigeria during the period January 2000 to December 2002. There were a total of 2,852 adult medical admissions during the study period out of which 575 were because of hypertension related morbidity. Only 302 (52.5%) of these were previously diagnosed as hypertensives). The most common hypertensive complication was cerebrovascular accident followed by congestive cardiac failure and chronic renal failure in order of decreasing frequency. The annual adult mortality rate from medical admissions was 5.6%. Mortality due to hypertensive complications constituted 10.5% of overall hospital adult mortality and 16.1% of deaths from medical causes. The annual mortality rate among HTA was 22.1% (252/1000) with a male:female ratio of 2:1. The contribution of systemic hypertension to adult morbidity and mortality is very significant. There is need for more concerted effort to create hypertension awareness, and to achieve good control and prevention of hypertensive complications.


Inhibition of terminal complement components in presensitized transplant recipients prevents antibody-mediated rejection leading to long-term graft survival and accommodation.
Ab-mediated rejection (AMR) remains the primary obstacle in presensitized patients following organ transplantation, as it is refractory to anti-T cell therapy and can lead to early graft loss. Complement plays an important role in the process of AMR. In the present study, a murine model was designed to mimic AMR in presensitized patients. This model was used to evaluate the effect of blocking the fifth complement component (C5) with an anti-C5 mAb on prevention of graft rejection. BALB/c recipients were presensitized with C3H donor skin grafts 7 days before heart transplantation from the same donor strain. Heart grafts, transplanted when circulating anti-donor IgG Abs were at peak levels, were rejected in 3 days. Graft rejection was characterized by microvascular thrombosis and extensive deposition of Ab and complement in the grafts, consistent with AMR. Anti-C5 administration completely blocked terminal complement activity and local C5 deposition, and in combination with cyclosporine and short-term cyclophosphamide treatment, it effectively prevented heart graft rejection. These recipients achieved permanent graft survival for >100 days with normal histology despite the presence of systemic and intragraft anti-donor Abs and complement, suggesting ongoing accommodation. Furthermore, double-transplant experiments demonstrated that immunological alterations in both the graft and the recipient were required for successful graft accommodation to occur. These data suggest that terminal complement blockade with a functionally blocking Ab represents a promising therapeutic approach to prevent AMR in presensitized recipients.


Right time - right location - right move: TRPs find motors for common functions.
TRP channels are localized at specialized sub-cellular compartments like filopodial tips, ciliary structures, growth cones and spines that have importance in the context of several sensory functions. Several motor proteins largely regulate these localizations. Recent studies indicate that both physical and genetic interactions exist between TRP channels with actin and microtubule-based motor proteins. These two groups of proteins share specialized and fine regulation underlying physiological functions. Indeed, mutations causing loss of these interactions and regulations result in development of pathophysiological disorders and syndromes. In this review we analyze the recent progress made in cell-biological, biochemical, electrophysiological and genetic studies and summarize the multi-dimensional crosstalk between TRP channels with different motor proteins.


Neuron-associated class III beta-tubulin isotype, microtubule-associated protein 2, and synaptophysin in human retinoblastomas in situ. Further immunohistochemical observations on the Flexner-Wintersteiner rosettes.
We studied by immunohistochemistry 26 retinoblastomas in situ using monoclonal antibodies specific for the neuron-associated class III beta-tubulin isotype (h beta 4), microtubule-associated protein 2 (MAP2), and synaptophysin. Anti-h beta 4 and anti-MAP2 immunostaining was consistently obtained in the Flexner-Wintersteiner rosettes, in fleurettes, in Homer Wright (neuroblastic) rosettes, and also variably among poorly differentiated tumor cells. A similar pattern of antisynaptophysin immunopositivity was seen, but was especially pronounced in the adluminal borders of cells forming the Flexner-Wintersteiner rosettes. The demonstration of h beta 4, MAP2, and synaptophysin epitopes in poorly differentiated and maturing neoplastic phenotypes in retinoblastomas attests to the neuronal character of this embryonal tumor. Immunoreactivity toward h beta 4 and MAP2 epitopes by poorly differentiated neoplastic cells may indicate early neuronal commitment in retinoblastoma. The consistent immunostaining of Flexner-Wintersteiner rosettes with monoclonal antibodies to h beta 4 and MAP2 is in keeping with the previous ultrastructural documentation of microtubules with a neuronal-like spatial organization present in the cells of these structures.


Alterations of physical and biochemical parameters of the R3327-CP rat prostate adenocarcinoma following hormonal manipulation of the host.
Several physical and biochemical parameters of a rapidly growing, hormonally responsive, poorly differentiated strain of Dunning R3327 rat prostatic adenocarcinoma (the CP strain) were monitored for 1 month during growth in control and hormonally manipulated male Fischer X Copenhagen rats. The tumor was implanted into control rats and into rats 1 month following orchiectomy. Twenty-nine days following tumor implantation, 1 group of unoperated rats was orchiectomized while the rats implanted subsequent to orchiectomy were repleted with pharmacological doses of testosterone. At 2 and 4 weeks following treatment, half the original number of rats from each group were sacrificed and the growth rate (doubling time), per cent of aneuploid cells and androgen receptor levels (total, cytoplasmic and nuclear) were determined for each tumor. Orchiectomy increased tumor doubling time, while testosterone repletion decreased it, demonstrating the hormonal dependence of this tumor strain. Orchiectomy also decreased the levels of aneuploid cells in the tumor; however, repletion of testosterone to rats orchiectomized prior to implantation did not restore the aneuploid cell number to control levels. A sensitive indicator of the hormonal status of the tumor was the per cent of androgen receptors in the nucleus. Tumors grown in rats orchiectomized after implantation had the lowest percentage of androgen receptor in the nucleus while orchiectomized rats repleted with testosterone had the highest percentage. Comparison of the levels of androgen receptors in the tumors from the various groups (androgen receptor per gram of tissue) unexpectedly revealed that tumors grown in the orchiectomized rats had slightly higher total receptor levels than did control tumors, while the tumors of orchiectomized rats repleted with testosterone had lower amounts than did the control tumors. In contrast to these findings, the prostates of orchiectomized rats replenished with testosterone had higher levels of total androgen receptor than did the prostates of control rats.


Aerosol deposition in health and disease.
The success of inhalation therapy is not only dependent upon the pharmacology of the drugs being inhaled but also upon the site and extent of deposition in the respiratory tract. This article reviews the main mechanisms affecting the transport and deposition of inhaled aerosol in the human lung. Aerosol deposition in both the healthy and diseased lung is described mainly based on the results of human studies using nonimaging techniques. This is followed by a discussion of the effect of flow regime on aerosol deposition. Finally, the link between therapeutic effects of inhaled drugs and their deposition pattern is briefly addressed. Data show that total lung deposition is a poor predictor of clinical outcome, and that regional deposition needs to be assessed to predict therapeutic effectiveness. Indeed, spatial distribution of deposited particles and, as a consequence, drug efficiency is strongly affected by particle size. Large particles (>6 μm) tend to mainly deposit in the upper airway, limiting the amount of drugs that can be delivered to the lung. Small particles (<2 μm) deposit mainly in the alveolar region and are probably the most apt to act systemically, whereas the particle in the size range 2-6 μm are be best suited to treat the central and small airways.


Hnrnpab regulates neural cell motility through transcription of Eps8.
Cell migration requires a complicated network of structural and regulatory proteins. Changes in cellular motility can impact migration as a result of cell-type or developmental stage regulated expression of critical motility genes. Hnrnpab is a conserved RNA-binding protein found as two isoforms produced by alternative splicing. Its expression is enriched in the subventricular zone (SVZ) and the rostral migratory stream within the brain, suggesting possible support of the migration of neural progenitor cells in this region. Here we show that the migration of cells from the SVZ of developing Hnrnpab-/- mouse brains is impaired. An RNA-seq analysis to identify Hnrnpab-dependent cell motility genes led us to Eps8, and in agreement with the change in cell motility, we show that Eps8 is decreased in Hnrnpab-/- SVZ tissue. We scrutinized the motility of Hnrnpab-/- cells and confirmed that the decreases in both cell motility and Eps8 are restored by ectopically coexpressing both alternatively spliced Hnrnpab isoforms, therefore these variants are surprisingly nonredundant for cell motility. Our results support a model where both Hnrnpab isoforms work in concert to regulate Eps8 transcription in the mouse SVZ to promote the normal migration of neural cells during CNS development.


Pharmacokinetic results on naproxen prodrugs based on poly(ethyleneglycol)s.
Five prodrugs of S(+)-2-(6-methoxy-2-naphthyl)propionic acid (naproxen), in which the drug was bound by ester linkages to diethyleneglycol (I), triethyleneglycol (II), octanediol (III), butyl-triethyleneglycol (IV), and butyl-tetraethyleneglycol (V), respectively, were prepared and tested for their pharmacokinetic properties after oral administration. It was found that bioavailabilities decreased in the order, and in all cases were lower than that of the free drug.


The epidemiology of sepsis: questioning our understanding of the role of race.
Race has been identified as an important risk factor for the development of sepsis and as a predictor of poor outcomes in sepsis. For example, black individuals have been demonstrated to be nearly twice as likely to develop sepsis and to have greater mortality from sepsis than white individuals. Recent data from a longitudinal cohort, which examined incident hospitalizations for infections occurring among participants in the Reasons for Geographic and Racial Differences in Stroke (REGARDS) cohort, contradicts this prior research. Investigators determined that black participants were significantly less likely than white participants to present to the hospital with either infection or sepsis. Although these results are intriguing, they highlight our inadequate understanding of the relationship between race and sepsis and motivate the need for higher quality epidemiologic research to isolate the true role of race in the development of sepsis.


Discovery of a series of dihydroquinoxalin-2(1H)-ones as selective BET inhibitors from a dual PLK1-BRD4 inhibitor.
Recent years have seen much effort to discover new chemotypes of BRD4 inhibitors. Interestingly, some kinase inhibitors have been demonstrated to be potent bromodomain inhibitors, especially the PLK1 inhibitor BI-2536 and the JAK2 inhibitor TG101209, which can bind to BRD4 with IC50 values of 0.025 μM and 0.13 μM, respectively. Although the concept of dual inhibition is intriguing, selective BRD4 inhibitors are preferred as they may diminish off-target effects and provide more flexibility in anticancer drug combination therapy. Inspired by BI-2536, we designed and prepared a series of dihydroquinoxalin-2(1H)-one derivatives as selective bromodomain inhibitors. We found compound 54 had slightly higher activity than (+)-JQ1 in the fluorescence anisotropy assay and potent antiproliferative cellular activity in the MM.1S cell line. We have successfully solved the cocrystal structure of 52 in complex with BRD4-BD1, providing a solid structural basis for the binding mode of compounds of this series. Compound 54 exhibited high selectivity over most non-BET subfamily members and did not show bioactivity towards the PLK1 kinase at 10 or 1 μM. From in vivo studies, compound 54 demonstrated a good PK profile, and the results from in vivo pharmacological studies clearly showed the efficacy of 54 in the mouse MM.1S xenograft model.


Comparison of Plantar Pressure Distribution During Walking After Two Different Surgical Treatments for Calcaneal Fracture.
The aim of our study was to compare gait in terms of foot loading and temporal variables after 2 different operative approaches (the extended lateral approach [ELA] and sinus tarsi approach). Twenty-two patients who sustained an intra-articular calcaneal fracture underwent plantar pressure distribution measurements 6 months after surgery. Measurements were performed while patients walked on the pedobarography platform. The values of dynamic variables were significantly lower on the operated limb in the ELA. In the sinus tarsi approach, no differences were observed between the operated and uninjured limbs (UIN) at peak pressure and at maximal vertical force. The values of temporal variables (contact time of the foot and of the heel) between the operated and UIN differed in the ELA. The hypothesis that differences in foot load between operated and UIN will be more significant in the ELA was confirmed. Our results showed that the differences in loading and temporal variables between the operated and the UIN persisted 6 months after surgery in both methods. The operated limb was less loaded, with the tendency to shift the load toward the midfoot and forefoot. After the less invasive sinus tarsi approach, the dynamic and temporal variables on the operated limb were nearly the same as those on the healthy one. The sinus tarsi surgical approach can be recommended for treatment of displaced calcaneal fractures.


Reversibility and "pH-T phase diagrams" of Rapana venosa hemocyanin and its structural subunits.
We have studied the stability and reassociation behaviour of native molecules of Rapana venosa hemocyanin and its two subunits, termed RvH1 and RvH2. In the presence of different concentrations of Ca(2+) and Mg(2+) ions and pH values, the subunits differ not only in their reassociation behaviour, but also in their formation of helical tubules and multidecamers. RvH1 revealed a greater stability at higher pH values compared to RvH2. Overall, the stability of reassociated RvH and its structural subunits was found to be pH-dependent. The increasing stability of native Hc and its subunits, shown by pH-induced CD transitions (acid and alkaline denaturation), can be explained with the formation of quaternary structure. The absence of a Cotton effect at temperatures 20-40 degrees C in the pH-transition curves of RvH2 indicates that this subunit is stabilized by additional "factors", e.g.: non-ionic/hydrophobic stabilization and interactions of carbohydrate moieties. A similar behaviour was observed for the T-transition curves in a wide pH interval for RvH and its structural subunits. At higher temperatures, many of the secondary structural elements are preserved especially at neutral pH, even at extreme high temperatures above 90 degrees C the protein structures resemble a "globule state".


Determination of sphingosine kinase activity in biological samples by liquid chromatography-tandem mass spectrometry.
Sphingosine kinase (SphK) is a key enzyme in modulating the levels of sphingosine 1-phosphate (S1P) as well as an important enzyme in numerous biological responses. Using C17-sphingosine as a substrate, we established a rapid, sensitive and highly efficient method for determination of SphK activity by analyzing the product C17-sphingosine 1-phosphate (C17-S1P) using liquid chromatography-tandem mass spectrometry. The standard curve for C17-S1P was linear over a wide range (10-1000 ng/mL) with correlation coefficient (r(2)) greater than 0.999. The lower limit of quantification for C17-S1P was 10 ng/mL. The K(m) values for C17-sphingosine and ATP were determined to be 28.17 and 188.5 microM, respectively. More importantly, the SphK activity dramatically increased in cultured HEK 293 cells expressing wild-type SphK1 as well as cells treated with tumor necrosis factor-alpha, a sphingosine kinase activator. In contrast, the SphK activity decreased in cultured HEK 293 cells treated with dimethylsphngosine, a sphingosine kinase inhibitor. In conclusion, this method was sensitive and rapid in the determination of SphK acitivity, providing striking utilities in exploring the sphingosine kinase signaling pathway and screening active compounds targeting SphK activity.


A small-molecule-based ternary data-storage device.
High-density data storage (HDDS) is urgently required to address ongoing rapid increases in amounts of information. Here we present a prototype sandwich device that has ternary data-storage performance. Three characteristic currents can be read out using a certain constant voltage after removal of the applied voltage. Two electron pull groups and one electron push group are identified as the species responsible for electron flow. We believe that our observation will offer an interesting and useful theoretical approach for a huge increase in the memory density of potential future devices.


Genetic influences on adult weight gain and maximum body mass index in male twins.
The National Heart, Lung, and Blood Institute Twin Study is a multicenter, longitudinal study of 514 white, male twin pairs examined during military induction at the mean age of 20 years (1943) and by the National Heart, Lung, and Blood Institute Twin Study at mean ages 48 (1971), 58 (1981), and 63 years (1986). Of these, 121 of 254 monozygotic pairs and 113 of 260 dizygotic pairs had complete data for all examinations. Using these data, genetic influences on maximum body mass index (BMI) and changes in BMI during this 43-year interval were estimated. BMI (kg/m2) was calculated for each examination and for the maximum weight as recalled by the participant at the last examination. Regression equations were fitted to each person's four-examination measurements to estimate the trend in weight change over adulthood. Twins gained an average of 0.11 kg/m2 per year from mean ages 20-63 years. Maximum BMI averaged 28.4 kg/m2 for monozygotic twins and 28.3 kg/m2 for dizygotic twins. The distribution of the ages at maximum BMI appeared to be bimodal for each zygosity, with modes around ages 20 and 60 years. Heritability (variation attributable to genetic factors) was estimated to be 0.71 (95% confidence interval 0.55-0.87) for maximum BMI and 0.70 (95% confidence interval 0.55-0.84) for trend in adult weight gain. In contrast, variability of BMI around the trend line showed no evidence of significant genetic determination.


Progranulin gene mutations associated with frontotemporal dementia and progressive non-fluent aphasia.
Frontotemporal lobar degeneration (FTLD) refers to a focal, non-Alzheimer form of cerebral degeneration that encompasses the distinct clinical syndromes of frontotemporal dementia (FTD), progressive non-fluent aphasia (PNFA) and semantic dementia. Some patients show tau-based pathological changes and in familial cases mutations have been identified in the microtubule-associated protein tau gene (MAPT) on chromosome 17q21. However, many cases are tau-negative, showing instead ubiquitin-immunoreactive (UBQ-ir) neuronal cytoplasmic inclusions and neurites, and in some familial cases UBQ-ir neuronal intranuclear inclusions of a lentiform appearance. Very recently, mutations have been identified in familial cases in the progranulin (PGRN) gene, also on chromosome 17q21. Clinical, pathological and molecular diversity within FTLD highlights the importance of careful examination of clinical-pathological-genetic relationships. This paper reports, for the first time, a clinico-pathological investigation of two FTLD families with PGRN mutations, and compares the clinical characteristics with those of patients studied in the department with MAPT mutations. The clinical profile associated with PGRN mutations constituted, in some patients, a prototypical picture of FTD and in others one of PNFA, both profiles occurring within the same family. Patients with PGRN mutations exhibited phonological deficits, whereas in patients with MAPT mutations language abnormalities, when present in addition to the prominent behavioural disorder, take the form of semantic disturbance. The findings provide compelling evidence for the link between FTD and PNFA, while raising the possibility of identifiable clinical differences between FTLD patients with MAPT and PGRN mutations.


[Operative treatment of ruptures of distal tendon of biceps muscle with a minimally invasive technique using suture anchors--clinical results].
Ruptures of the distal tendon of the biceps muscle should be treated operatively. A minimally invasive technique and the use of suture anchors may decrease the intraoperative risks. But can satisfactory clinical results be achieved? We present the results of a clinical study. Between 1999 and 2005 nine patients were operated for distal biceps tendon avulsion. A minimally invasive operating technique using a suture anchor was established and the technique is described. Rantanen and Orava's score was used to compare clinical and radiological findings. Clinical follow-up was performed at 6 months after operation. In six patients excellent results according to Rantanen and Orava were achieved. No loss of strength was noticed in 6 of 9 cases. Range of motion was unlimited in six patients. In two cases we found heterotopic ossifications. On the basis of our results we suggest that minimally invasive surgery using a suture anchor should be performed more often. The technique described leads to results that are comparable to those found in the literature.


Two-stage treatment with preoperative endoscopic retrograde cholangiopancreatography (ERCP) compared with single-stage treatment with intraoperative ERCP for patients with symptomatic cholelithiasis with possible choledocholithiasis.
An ideal treatment for choledocholithiasis in the laparoscopic era has not been established. The objective of this study was to elucidate whether a treatment strategy of performing intraoperative endoscopic retrograde cholangiopancreatography (ERCP) during laparascopic cholecystectomy (when choledocholithiasis is confirmed by intraoperative cholangiography) is better for patients with suspected common bile duct stones than the current strategy (preoperative ERCP followed by laparoscopic cholecystectomy). This was a prospective randomized study to evaluate which of these two approaches was most benefit- and cost-effective for patients with intermediate risk of choledocholithiasis. Patients underwent either preoperative ERCP followed by a laparoscopic cholecystectomy a few weeks later (the "preoperative ERCP" group) or intraoperative ERCP (the "intraoperative ERCP" group). Intraoperative ERCP was performed using the rendezvous technique. There were 64 patients in the preoperative ERCP group and 59 patients in the intraoperative ERCP group. The demographic and clinical characteristics of the two groups were similar, except that the bilirubin and gamma-glutamyl transferase (GGT) levels and the number of patients treated on an inpatient basis were higher in the preoperative ERCP group. Success rates were similar (96.6 % in the preoperative ERCP group vs. 90.2 % in the intraoperative ERCP group in the per-protocol study). Total morbidity, post-ERCP morbidity, and post-ERCP acute pancreatitis rates were higher in the preoperative ERCP group, but there were no differences between the two groups in the frequency of residual common bile duct stones, the conversion rate to open cholecystectomy, or surgical morbidity. The length of hospital stay and costs were lower in the intraoperative ERCP group despite the longer surgical times in this group. Univariate analysis did not find any relationship between morbidity and total bilirubin or GGT. Logistic regression analysis confirmed that morbidity was related only to the treatment group and the time spent in the operating room: the relative risk (RR) was 4.37 for morbidity and 1.015 for the time spent in the operating room); the RR for papillotomy was 5.49. Both treatment approaches were equally effective but the intraoperative ERCP group had less morbidity, a shorter hospital stay, and reduced costs. The lower morbidity in the intraoperative ERCP group resulted from the lower rate of papillotomy and lower rates of post-ERCP pancreatitis and cholecystitis. Total morbidity was principally related to the type of treatment approach used.


Dietary induction of sudden death syndrome in broiler breeders.
Sudden death syndrome in broiler breeders at the point of lay was a serious problem in Australia for 18 months. This condition was reproduced by a combination of dietary and management practices. Chickens fed a diet low in potassium, phosphorus, protein, and energy had plasma potassium and phosphorus levels significantly lower than the levels seen in the controls. The plasma values of the test breeders were similar to levels seen in field cases. There was a significant difference (P less than 0.01) in the venous acid-base balance between the test chickens and controls. Test chickens were alkalotic and had a 6% mortality rate between 18 and 30 weeks of age.


Double blind, randomized, placebo-controlled study of dexamethasone therapy for hematogenous septic arthritis in children.
Septic arthritis is associated with residual dysfunction in 10 to 25% of affected children. Concentrations of cytokines detected in synovial fluid of children with bacterial arthritis correlate with the severity of inflammation. Treatment with dexamethasone decreased cartilage degradation in experimental Haemophilus influenzae b and Staphylococcus aureus arthritis. To decrease the number of patients with residual dysfunction of the affected joint at the end of therapy and at 6 and 12 months and to speed clinical recovery by the administration of dexamethasone. In a double blind manner we randomly selected 123 children with suspected hematogenous bacterial arthritis to receive dexamethasone or saline for 4 days. Antibiotic therapy was tailored according to age and the recovered pathogen. Of the 123 children enrolled, 61 were assigned to the dexamethasone group and 62 to the placebo group. Only 50 and 50 patients in each group were evaluable. The 2 groups of patients were comparable with respect to age, sex, duration of symptoms, pathogen, affected joint and therapeutic and diagnostic procedures. Staphylococcus aureus accounted for 67% of the isolates, Haemophilus influenzae type b for 13% and Streptococcus pneumoniae for 9%. Dexamethasone therapy reduced residual dysfunction at the end of therapy, P = 0.000068; at 6 months, P = 0.00007; and at 12 months, P = 0.00053 of follow-up and shortened the duration of symptoms (P = 0.001) during the acute phase. The 26% incidence of residual dysfunction in the control patients was similar to the 25% found in other series. A short course of dexamethasone reduced residual joint dysfunction and shortened significantly the duration of symptoms in children with documented hematogenous septic arthritis. These results suggest that a 4-day course of low dose dexamethasone given early benefits children with hematogenous septic arthritis.


Safe and effective use of transcutaneous blood gas monitors.
The Medical Devices Amendments of 1976 (PL 94-295) will compel users of transcutaneous monitors to comply with Class II, Performance Standards. These standards will have to provide reasonable assurance of the safety and effectiveness of the device. Similar standards are in preparation for other cardiac and respiratory monitors. Dangers of tcPO2 monitors include shock and burn, which requires attention to the insulation of the monitor from the power line and from recorders used to record the output. Optimal safe time vs. temperature limits have not yet used to record to output. Optimal safe time vs. temperature limits have not yet been established, and these may vary with age, skin site, blood pressure and body and environmental temperature. The usefulness of alarms for PO2, sensor temperature and heating power, and their limits remain ot be defined by extensive clinical use. Standards for drift, reproducibility, recalibration interval and accuracy will have to be set after more extensive clinical use especially by groups not involved in the initial prototype development and testing. In order to prove that these devices are also effective, physicians will have to document that care would have been different, and might have impaired patient safety, if the device had not been used.


Elevated gene expression levels distinguish human from non-human primate brains.
Little is known about how the human brain differs from that of our closest relatives. To investigate the genetic basis of human specializations in brain organization and cognition, we compared gene expression profiles for the cerebral cortex of humans, chimpanzees, and rhesus macaques by using several independent techniques. We identified 169 genes that exhibited expression differences between human and chimpanzee cortex, and 91 were ascribed to the human lineage by using macaques as an outgroup. Surprisingly, most differences between the brains of humans and non-human primates involved up-regulation, with approximately 90% of the genes being more highly expressed in humans. By contrast, in the comparison of human and chimpanzee heart and liver, the numbers of up- and down-regulated genes were nearly identical. Our results indicate that the human brain displays a distinctive pattern of gene expression relative to non-human primates, with higher expression levels for many genes belonging to a wide variety of functional classes. The increased expression of these genes could provide the basis for extensive modifications of cerebral physiology and function in humans and suggests that the human brain is characterized by elevated levels of neuronal activity.


Transient Hypoglycemia and Biochemical Differences in Infants Less Than 1,250 G at Birth Fed Human Milk with Human Milk-Derived Fortifier versus Cow Milk-Derived Fortifier.
Fortification of human milk (HM) with either human milk-derived fortifier (HMDF) or cow milk-derived fortifier (CMDF) is important in preterm infants. The objective is to compare the incidence of hypoglycemia, and biochemical values in infants less than 1,250 g at birth fed HMDF versus CMDF. It is a retrospective cohort study on infants less than 1,250 g at birth who were fed with HMDF or CMDF. Hypoglycemia was defined as blood glucose (BG) level equal to or less than 60 mg/dL within 72 hours of full enteral feeds when off total parenteral nutrition and intravenous fluids. Ninety infants were enrolled (HMDF = 61, CMDF = 29). HMDF group had a higher rate of hypoglycemia (46 vs. 24%; p = 0.048) after achievement of full enteral feeding. The median minimum BG was lower (61 vs. 71; p ≤ 0.01), while blood urea nitrogen (12 vs. 6; p ≤ 0.01) and albumin (3.1 vs. 2.7; p ≤ 0.01) were higher in HMDF group compared with CMDF. At full enteral feedings in infants less than 1,250 g at birth, an HMDF diet may predispose to hypoglycemia needing intervention. Close monitoring of BG levels once off parenteral nutrition is recommended. · Exclusive human milk (EHM) feeding results in better nutritional indices.. · EHM feeding at higher calorie/ounce improves growth.. · Blood glucose needs to be monitored when off TPN during EHM feeding..


Persistence of Anti-S1 IgG against SARS-CoV-2 Eight Months after the Booster Dose of Vaccine in Naive and Previously Infected Healthcare Workers.
Our aim was to evaluate the immune response of healthcare workers included in the RIPOVAC study, after receiving a booster dose (third dose), in terms of intensity and persistence of induced antibodies. In the second phase of the RIPOVAC study, between December 2021 and January 2022, eight months after the second dose, 389 voluntary, immunocompetent, non-pregnant healthcare workers received a booster dose of SARS-CoV-2 vaccine, and a serum sample was obtained. Two groups of patients were established: with and without previous SARS-CoV-2 infection. In order to quantify anti-S1 IgG (AU/mL) we used CMIA (Abbott). All of the health workers were anti-S IgG positive 8 months after receiving the booster dose of the vaccine, with a mean of 17,040 AU/mL. In 53 patients without previous infection, antibody levels increased by a mean of 10,762 AU/mL. This figure is seven times higher than the one produced after the second dose (1506 AU/mL). The booster dose produces a robust elevation of the antibody level, which persists at 8 months, with levels significantly higher than those reached after the second dose, which allow one to predict a persistence of more than one year. The study demonstrates the efficacy of the booster dose of anti-SARS-CoV-2 vaccines.


Prospective relations between rejection and depression in young adolescents.
The purpose of this longitudinal study was to examine the relations between rejection and depression across 3 years in young adolescents who varied with regard to their risk for depression. The sample consisted of 240 adolescents who were assessed in grades 6, 7, and 8. The assessment of rejection was based on adolescent-, mother-, and teacher-report, and depression assessment was based on adolescent- and mother-report and clinician ratings. Structural equation modeling indicated that rejection prospectively predicted depression. The authors did not find that depression prospectively predicted rejection, but such a relation cannot be ruled out because of strong cross-sectional correlations between depression and rejection.


Extracorporeal life support for severe acute respiratory distress syndrome.
To provide a summary of the recent literature on extracorporeal membrane oxygenation (ECMO) in adults with severe acute respiratory distress syndrome (ARDS), focusing on advances in equipment, current conventional and unconventional indications, complications, and future applications. ECMO use has increased during the past 5 years. Advances in cannulation, circuit design, and patient selection have made it a safer therapeutic option in severe ARDS, and its use has become more widespread for nonconventional indications. High-quality evidence for the routine use of ECMO for management of adult patients with severe ARDS is still lacking. An ongoing randomized controlled trial (ECMO to rescue lung injury in severe ARDS) will contribute valuable data to guide clinical decisions to opt for this supportive therapy.


Measurement of 238U, 232Th, 222Rn and 220Rn Contents in Optical Contact Lenses: Resulting Alpha and Beta Equivalent Doses to the Eye Tissues of Adult Patients.
To estimate equivalent doses received by eye tissues of patients, contents of uranium (238U), thorium (232Th), radon (222Rn), and thoron (220Rn) were determined inside various optical contact lenses used for vision correction. 238U, 232Th, 222Rn, and 220Rn contents varied between (3.44 ± 0.24) mBq kg-1 and (18.3 ± 1.28) mBq kg-1, (0.57 ± 0.04) mBq kg-1 and (3.53 ± 0.25) mBq kg-1, (3.44 ± 0.24) mBq kg-1 and (18.3 ± 1.28) mBq kg-1, and (0.57 ± 0.04) mBq kg-1 and (3.53 ± 0.25) mBq kg-1, respectively. New external dosimetric models, depending on the cornea eye surface of patients, 238U, 232Th, and 222Rn concentrations inside optical contact lenses, half-life of the emitting radionuclides, and exposure time of patients, have been developed. It has been shown that alpha-particles emitted by the 238U and 232Th series inside the studied optical lenses transfer their energies essentially to the cornea tissues whereas the emitted beta-particles may reach and lose their energies in the crystalline lens of eyes of patients. Alpha-equivalent doses received by eye tissues of patients due to the diffusion of 222Rn and 220Rn gases present in the considered optical lenses were determined. The higher value of the total (alpha plus beta) equivalent dose to the left and right eyes of adult patients wearing optical contact lenses (14 hours per day) has been found equal to 1.32 mSv y-1 cm - 2. It is recommended for patients to reduce the wearing period of optical contact lenses to reduce eye disease risks such as cataract.


Pituitary Adenoma With Apoplexy Presenting As Unilateral Third Nerve Palsy.
Pituitary adenomas are one of the most common intracranial tumors. Non-functioning macroadenomas are usually diagnosed when they cause symptoms due to the mass effect on surrounding structures. We present the case of a 48-year-old man who presented with a headache associated with ptosis of the right eye and right-sided blurry vision for three days. Initial computerized tomography (CT) scan of the head did not report a mass, acute infarct, or hemorrhage. He was given 325mg of aspirin for concern of a stroke while waiting for magnetic resonance imaging (MRI) of the brain, which was done the next day and revealed a pituitary macroadenoma with hemorrhage, mass effect, and compression of the optic chiasm consistent with pituitary apoplexy. He ultimately underwent trans-sphenoidal resection of the tumor; however, his surgery was delayed for five days as he had received a high dose of aspirin in the Emergency Room. His adrenocorticotropic hormone (ACTH), cortisol, and testosterone levels were found to be quite low. He was administered stress dose steroids peri-operatively and ultimately discharged on indefinite hydrocortisone therapy and endocrinology follow-up. Our case highlights a serious complication of pituitary adenomas that can occur called pituitary apoplexy which is caused by acute ischemic infarction or hemorrhage in the pituitary. It needs prompt identification and management. Our case also emphasizes the importance of recognizing pituitary apoplexy as one of the causes of sudden onset cranial nerve deficits, as it is a rare presentation of pituitary adenomas that can be taken for a stroke in the Emergency Department.


[Osteoarticular manifestations during liver and gastrointestinal diseases].
The most important osteoarticular complications occurring during gastrointestinal and liver diseases are reviewed. Although etiopathogenesis of these disorders is not yet fully understood, their knowledge is required to gastroenterologists for two main reasons: to establish a correct diagnosis and to start an appropriate therapy.


The communities of terrestrial macrofungi in different forest types in vicinities of Khanty-Mansiysk (middle taiga zone of West Siberia).
The diversity of macrofungi in the vicinities of Khanty-Mansiysk (Yugra, Russia) was surveyed using a method of permanent sampling plots. Ten plots, each consisting of a number of micro-plots, were established in several different communities ranging from old-growth mixed taiga forest to its derivatives in cutting succession and bogged areas. For more complete registration of the mycota, plots were supplemented with random walking routes directly nearby. Survey results were subjected to various quantitative analyses which allowed not only to evaluate the diversity of fungi but also to obtain valuable information on occurrence, abundance and ecology of individual species as well as community structure and its dynamics in the course of ecological succession. The paper reports the results of the first year of observations. 460 species of terrestrial macrofungi revealed in a poorly explored area in middle taiga of West Siberia. The plot-based study revealed differences between communities of terrestrial macrofungi of old coniferous forests, their after-cutting secondary formations and bogged stages. The survey allowed to reveal records of 3 species listed in the Red Data Book of Russia and 9 species listed in the Red Data Book of Yugra.


Nucleotide sequence structure and consistency of a developmentally regulated DNA deletion in Tetrahymena thermophila.
DNA deletion by site-specific chromosome breakage and rejoining occurs extensively during macronuclear development in the ciliate Tetrahymena thermophila. We have sequenced both the micronuclear (germ line) and rearranged macronuclear (somatic) forms of one region from which 1.1 kilobases of micronuclear DNA are reproducibly deleted during macronuclear development. The deletion junctions lie within a pair of 6-base-pair direct repeats. The termini of the deleted sequence are not inverted repeats. The precision of deletion at the nucleotide level was also characterized by hybridization with a synthetic oligonucleotide matching the determined macronuclear (rejoined) junction sequence. This deletion occurs in a remarkably sequence-specific manner. However, a very minor degree of variability in the macronuclear junction sequences was detected and was shown to be inherent in the mechanism of deletion itself. These results suggest that DNA deletion during macronuclear development in T. thermophila may constitute a novel type of DNA recombination and that it can create sequence heterogeneity on the order of a few base pairs at rejoining junctions.


Design, Fabrication, and Accuracy of a Novel Noncovering Lock-Mechanism Bilateral Patient-Specific Drill Guide Template for Nondeformed and Deformed Thoracic Spines.
Background: Pedicle screw (PS) placement has been widely used in fusion surgeries on the thoracic spine. Achieving cost-effective yet accurate placements through nonradiation techniques remains challenging. Questions/Purposes: Novel noncovering lock-mechanism bilateral vertebra-specific drill guides for PS placement were designed/fabricated, and their accuracy for both nondeformed and deformed thoracic spines was tested. Methods: One nondeformed and 1 severe scoliosis human thoracic spine underwent computed tomographic (CT) scanning, and 2 identical proportions of each were 3-dimensional (3D) printed. Pedicle-specific optimal (no perforation) drilling trajectories were determined on the CT images based on the entry point/orientation/diameter/length of each PS. Vertebra-specific templates were designed and 3D printed, assuring minimal yet firm contacts with the vertebrae through a noncovering lock mechanism. One model of each patient was drilled using the freehand and one using the template guides (96 pedicle drillings). Postoperative CT scans from the models with the inserted PSs were obtained and superimposed on the preoperative planned models to evaluate deviations of the PSs. Results: All templates fitted their corresponding vertebra during the simulated operations. As compared with the freehand approach, PS placement deviations from their preplanned positions were significantly reduced: for the nonscoliosis model, from 2.4 to 0.9 mm for the entry point, 5.0° to 3.3° for the transverse plane angle, 7.1° to 2.2° for the sagittal plane angle, and 8.5° to 4.1° for the 3D angle, improving the success rate from 71.7% to 93.5%. Conclusions: These guides are valuable, as the accurate PS trajectory could be customized preoperatively to match the patients' unique anatomy. In vivo studies will be required to validate this approach.


Exploring Chinese Women's Perception of Cervical Cancer Risk as It Impacts Screening Behavior: A Qualitative Study.
Chinese women rarely undergo regular cervical screening. Women's decisions about cervical screening may be influenced by their perception of the risk of cervical cancer. Therefore, understanding how women perceive the risk of cervical cancer, how personal risk factors are interpreted, and the influence of cultural issues on cervical screening behavior is important. The aim of this study is to understand cervical cancer risk perception and the role of personal risk factors as they influence screening behavior among Chinese women. An exploratory qualitative research design was used. We conducted semistructured, in-depth interviews of 27 women in Changsha, a medium-size city in Hunan province, China. Participants identified that cervical cancer had serious consequences, but they distanced themselves psychologically from the disease because they felt that "cervical cancer is a shameful and deadly disease." Although women identified some of the risk factors for the disease, they had little specific knowledge of human papillomavirus infection, its association with cervical cancer, and the importance of cervical screening. This study contributes new knowledge to the understanding of cervical screening behavior within a specific social and cultural context. Better efforts should be made to educate Chinese women on the risk of cervical cancer and emphasize that effective cancer treatments are available and that there is a good chance of survival. Nurses working with Chinese women should ensure that the information they provide is culturally sensitive, particularly by acknowledging the normative beliefs of this population group.


Introduction: surgery of the central skull base.
With his anatomical studies of the parasellar space, the so-called cavernous sinus (CS), Taptas opened Pandora's box more than 60 years ago. Parkinson continued the anatomical studies, and operated on vascular lesions in the CS with the help of extracorporeal circulation. The need for endovascular treatment of intracavernous internal carotid artery (ICA) aneurysms, as well as carotid-cavernous fistulas (CCFs), was obvious. Serbinenko started with the endovascular treatment of CCFs and ICA aneurysms using a balloon. At nearly the same time, Hakuba undertook surgical treatment of tumorous lesions in the region. Glascock studied the ICA in relation to the petrous bone, and with his studies of the ICA and this artery's relationship to the other structures, it became clear that further understanding of the pathological entities in the parasellar space hinged on additional microanatomical studies.


Influence of microbial stimulation on hypergammaglobulinemia and autoantibody production in pristane-induced lupus.
Pristane induces a lupus-like syndrome characterized by autoantibody production and glomerulonephritis in nonautoimmune strains of mice. Although it has been suggested that this syndrome results from nonspecific immune activation, there is little evidence so far that B cells are activated nonspecifically by pristane or that this promotes autoimmunity. In this study, we examined whether polyclonal hypergammaglobulinemia occurs in pristane-induced lupus, and its relationship to the production of anti-DNA, nRNP/Sm, and Su autoantibodies. In conventionally housed mice, there was a marked increase in total IgM and IgG3 2 weeks after i.p. pristane injection, followed by increased IgG1, IgG2a, and IgG2b levels. IgM levels were higher in pristane-treated specific pathogen-free (SPF) mice than in conventionally housed mice, whereas IgG and IgA levels were reduced. Pristane induced anti-nRNP/Sm and Su autoantibodies in SPF mice, but their onset was delayed and levels were lower than those in conventionally housed mice. There was no consistent relationship between total IgG1, 2a, and 2b hypergammaglobulinemia and production of anti-nRNP/Sm and Su autoantibodies. Moreover, the total Ig levels were similar in the anti-nRNP/Sm-positive and -negative groups. In contrast, production of IgM anti-ssDNA antibodies paralleled IgM hypergammaglobulinemia in some, but not all, mice. These studies indicate that pristane-induced lupus is associated with marked hypergammaglobulinemia, the magnitude of which is influenced by the microbial environment. However, anti-nRNP/Sm and Su autoantibody production is at least partly independent of polyclonal B cell activation. The data strongly suggest that pristane-induced lupus is not exclusively the consequence of nonspecific immune stimulation. They also point to the importance of microbial stimulation in the development of hypergammaglobulinemia in this inducible lupus model.


Application of nanodiagnostics in point-of-care tests for infectious diseases.
Although tremendous efforts have been put into the treatment of infectious diseases to prevent epidemics and mortality, it is still one of the major health care issues that have a profound impact on humankind. Therefore, the development of specific, sensitive, accurate, rapid, low-cost, and easy-to-use diagnostic tools is still in urgent demand. Nanodiagnostics, defined as the application of nanotechnology to medical diagnostics, can offer many unique opportunities for more successful and efficient diagnosis and treatment for infectious diseases. In this review, we provide an overview of the nanodiagnostics for infectious diseases from nanoparticle-based, nanodevice-based, and point-of-care test (POCT) platforms. Most importantly, emphasis focused on the recent trends in the nanotechnology-based POCT system. The current state-of-the-art and most promising point-of-care nanodiagnostic technologies, including miniaturized diagnostic magnetic resonance platform, magnetic barcode assay system, cell phone-based polarized light microscopy platform, cell phone-based dongle platform, and paper-based POCT platform, for infectious diseases were fully examined. The limitations, challenges, and future trends of the nanodiagnostics in POCTs for infectious diseases are also discussed.


Administration of auditory stimulation during recovery after REM sleep deprivation.
Rapid eye movement (REM) sleep deprivation and auditory stimulation (ADS), separately, increase REM sleep in rats, cats and humans. The main goal of the present study was to test whether administration of ADS during REM sleep rebound has a synergistic effect on REM sleep elicitation. Male Wistar rats were implanted with standard sleep recording electrodes. Following the recovery period, animals were randomly assigned to the following conditions: undeprived (i.e. control) and 24, 48, 96 and 120 hours of REM sleep deprivation by the platform method. Undeprived and REM sleep-deprived animals were divided into two groups, with and without ADS. ADS was a "beep" of 80 dB and 2,000 Hz, lasting 20 msec every 10 seconds. This stimulus was applied for the first 4 hours of sleep recordings after deprivation. After that, animals were recorded for another 4 hours. In the undeprived situation, the group that received ADS increased REM sleep approximately 70% above the group that did not receive ADS, as has been reported previously (REM sleep without ADS: 38.1 +/- 13.84 vs. with ADS: 64.6 +/- 11.8, p < 0.005). No synergistic effect was observed between REM sleep deprivation and ADS for any REM sleep-deprivation schedule. This result may be explained as an increase in the excitability pattern of pontine neurons and/or changes in the cholinergic system due to REM sleep deprivation that could not be further increased by ADS.


Asymmetric exponential amplification reaction on a toehold/biotin featured template: an ultrasensitive and specific strategy for isothermal microRNAs analysis.
The sensitive and specific analysis of microRNAs (miRNAs) without using a thermal cycler instrument is significant and would greatly facilitate biological research and disease diagnostics. Although exponential amplification reaction (EXPAR) is the most attractive strategy for the isothermal analysis of miRNAs, its intrinsic limitations of detection efficiency and inevitable non-specific amplification critically restrict its use in analytical sensitivity and specificity. Here, we present a novel asymmetric EXPAR based on a new biotin/toehold featured template. A biotin tag was used to reduce the melting temperature of the primer/template duplex at the 5' terminus of the template, and a toehold exchange structure acted as a filter to suppress the non-specific trigger of EXPAR. The asymmetric EXPAR exhibited great improvements in amplification efficiency and specificity as well as a dramatic extension of dynamic range. The limit of detection for the let-7a analysis was decreased to 6.02 copies (0.01 zmol), and the dynamic range was extended to 10 orders of magnitude. The strategy enabled the sensitive and accurate analysis of let-7a miRNA in human cancer tissues with clearly better precision than both standard EXPAR and RT-qPCR. Asymmetric EXPAR is expected to have an important impact on the development of simple and rapid molecular diagnostic applications for short oligonucleotides.


Use of EHRs data for clinical research: Historical progress and current applications.
The benefits of reusing EHR data for clinical research studies are numerous. They portend the opportunity to bring new therapies to patients sooner, potentially at a lower cost, and to accelerate learning health cycles-through faster data acquisition in clinical research studies. Metrics have proven that time can be saved, workflow and processes streamlined, and data quality increased significantly. Pilot projects and now actual investigational trials used for regulatory submissions have shown that these benefits support the transformation of clinical research by leveraging EHRs for research. Panelists at a recent collaborative focused on bridging clinical research and clinical care offered varying perspectives on how the latest standards and technologies could be leveraged to facilitate data transfer from EHR systems into clinical research databases, as well as the associated improvements in data quality. Panelists also discussed other avenues to leverage EHR in clinical research. Improvements and exciting possibilities notwithstanding, much work remains. Data ownership and access, attention to metadata and structured data for data sharing, and broader adoption of global standards are key areas for collaboration. With the steady increase in adoption of EHRs around the world, this is an excellent time for all stakeholders to work together and create an environment such that EHRs can be used more readily for research. The capacity for research can thus be increased to provide more high-quality information that will contribute to rapid continuous learning health systems from which all patients can benefit.


Lie construction affects information storage under high memory load condition.
Previous studies indicate that lying consumes cognitive resources, especially working memory (WM) resources. Considering the dual functions that WM might play in lying: holding the truth-related information and turning the truth into lies, the present study examined the relationship between the information storage and processing in the lie construction. To achieve that goal, a deception task based on the old/new recognition paradigm was designed, which could manipulate two levels of WM load (low-load task using 4 items and high-load task using 6 items) during the deception process. The analyses based on the amplitude of the contralateral delay activity (CDA), a proved index of the number of representations being held in WM, showed that the CDA amplitude was lower in the deception process than that in the truth telling process under the high-load condition. In contrast, under the low-load condition, no CDA difference was found between the deception and truth telling processes. Therefore, we deduced that the lie construction and information storage compete for WM resources; when the available WM resources cannot meet this cognitive demand, the WM resources occupied by the information storage would be consumed by the lie construction.


Gene-expression Profiling in Non-small Cell Lung Cancer with Invasion of Mediastinal Lymph Nodes for Prognosis Evaluation.
The aim of the study was to determine the pathways and expression profile of the genes that might predict response to neoadjuvant chemotherapy in patients with stage IIIA non-small cell lung cancer (NSCLC). We evaluated, by microarray, the gene-expression profile of tumoral mediastinal lymph node samples surgically removed from 27 patients with stage IIIA NSCLC before neoadjuvant chemotherapy treatment. Depending on the response to the induction treatment, the patients were divided in two groups: group A: patients whose disease evolved, stabilized or who had minor response to chemotherapy, and group B: patients whose disease stabilized or had major response to chemotherapy. The microarray experiments identified 1,127 genes with a modified expression in the tumoral tissue compared to normal tissue with p≤0.05 and 44 genes with p≤0.01. The identified up-regulated genes between tumoral versus normal tissue included collagen, type I, alpha 1 (COL1A1), inhibin beta A (INHBA) and thioredoxin interacting protein (TXNIP). Pathways identified with a false-discovery rate of <0.005 included: cytokine pathways, focal adhesion or extracellular matrix receptor interaction. Our approach identified important characteristics of NSCLC and pointed-out molecular differences between sub-groups of patients based on their response to therapy.


Modulation of mitochondrial metabolism as a biochemical trait in blood feeding organisms: the redox vampire hypothesis redux.
Hematophagous organisms undergo remarkable metabolic changes during the blood digestion process, increasing fermentative glucose metabolism, and reducing respiratory rates, both consequence of functional mitochondrial remodeling. Here, we review the pathways involved in energy metabolism and mitochondrial functionality in a comparative framework across different hematophagous species, and consider how these processes regulate redox homeostasis during blood digestion. The trend across distinct species indicate that a switch in energy metabolism might represent an important defensive mechanism to avoid the potential harmful interaction of oxidants generated from aerobic energy metabolism with products derived from blood digestion. Indeed, in insect vectors, blood feeding transiently reduces respiratory rates and oxidant production, irrespective of tissue and insect model. On the other hand, a different scenario is observed in several unrelated parasite species when exposed to blood digestion products, as respiratory rates reduce and mitochondrial oxidant production increase. The emerging picture indicates that re-wiring of energy metabolism, through reduced mitochondrial function, culminates in improved tolerance to redox insults and seems to represent a key step for hematophagous organisms to cope with the overwhelming and potentially toxic blood meal.


Temporal mTOR inhibition protects Fbxw7-deficient mice from radiation-induced tumor development.
FBXW7 acts as a tumor suppressor in numerous types of human cancers through ubiquitination of different oncoproteins including mTOR. However, how the mutation/loss of Fbxw7 results in tumor development remains largely unknown. Here we report that downregulation of mTOR by radiation is Fbxw7-dependent, and short-term mTOR inhibition by rapamycin after exposure to radiation significantly postpones tumor development in Fbxw7/p53 double heterozygous (Fbxw7+/-p53+/-) mice but not in p53 single heterozygous (p53+/-) mice. Tumor latency of rapamycin treated Fbxw7+/-p53+/- mice is remarkably similar to those of p53+/- mice while placebo treatedFbxw7+/-p53+/- mice develop tumor significantly earlier than placebo treated p53+/- mice. Furthermore, we surprisingly find that, although temporal treatment of rapamycin is given at a young age, the inhibition of mTOR activity sustainably remains in tumors. These results indicate that inhibition of mTOR signaling pathway suppresses the contribution of Fbxw7 loss toward tumor development.


Emerging technologies for the clinical microbiology laboratory.
In this review we examine the literature related to emerging technologies that will help to reshape the clinical microbiology laboratory. These topics include nucleic acid amplification tests such as isothermal and point-of-care molecular diagnostics, multiplexed panels for syndromic diagnosis, digital PCR, next-generation sequencing, and automation of molecular tests. We also review matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) and electrospray ionization (ESI) mass spectrometry methods and their role in identification of microorganisms. Lastly, we review the shift to liquid-based microbiology and the integration of partial and full laboratory automation that are beginning to impact the clinical microbiology laboratory.