Cumulative and acute toxicity of repeated high-dose tobramycin treatment in cystic fibrosis.
Forty-six patients with cystic fibrosis and chronic bronchopulmonary Pseudomonas aeruginosa infection entered a study of tobramycin-related chronic and acute nephro- and acousticovestibular toxicity. The patients (mean age, 15.7 years) had previously received 2-week courses of tobramycin therapy, for a mean cumulative total of 279 days each. The cumulative tobramycin dose ranged from 632 to 7,644 mg/kg. The patients were studied before and at the end of a 2-week course of treatment with tobramycin (10 to 20 mg/kg per day) to discriminate between acute and chronic toxicity. In patients studied at the beginning of the present course of treatment, the glomerular filtration rate, measured as 24-h creatinine clearance, did not correlate with the cumulative dose of tobramycin received during previous courses. Eighteen patients (39%) had a reduced glomerular filtration rate compared with normal values (mean, 12.5% reduction) but normal serum creatinine values. Two patients (5%) had a high-frequency hearing deficit (above 8 kHz), but only one deficit was possibly related to tobramycin. No chronic vestibular toxicity was observed. During the course of treatment, no patients developed acute nephrotoxicity. After 2 weeks of treatment 32% had a slightly reduced hearing threshold (15 to 30 dB) in two or more high frequencies, and 28% had a fall in vestibular response greater than 25% of the initial value but remained within normal limits. Thus, the acute and chronic toxicity of repeated high-dose tobramycin treatment in cystic fibrosis patients seems to be very mild.


Recognition of Histone H3 Methylation States by the PHD1 Domain of Histone Demethylase KDM5A.
PHD reader domains are chromatin binding modules often responsible for the recruitment of large protein complexes that contain histone modifying enzymes, chromatin remodelers, and DNA repair machinery. A majority of PHD domains recognize N-terminal residues of histone H3 and are sensitive to the methylation state of Lys4 in histone H3 (H3K4). Histone demethylase KDM5A, an epigenetic eraser enzyme that contains three PHD domains, is often overexpressed in various cancers, and its demethylation activity is allosterically enhanced when its PHD1 domain is bound to the H3 tail. The allosteric regulatory function of PHD1 expands roles of reader domains, suggesting unique features of this chromatin interacting module. Our previous studies determined the H3 binding site of PHD1, although it remains unclear how the H3 tail interacts with the N-terminal residues of PHD1 and how PHD1 discriminates against H3 tails with varying degrees of H3K4 methylation. Here, we have determined the solution structure of apo and H3 bound PHD1. We observe conformational changes occurring in PHD1 in order to accommodate H3, which interestingly binds in a helical conformation. We also observe differential interactions of binding residues with differently methylated H3K4 peptides (me0, me1, me2, or me3), providing a rationale for PHD1's preference for lower methylation states of H3K4. We further assessed the contributions of various H3 interacting residues in the PHD1 domain to the binding of H3 peptides. The structural details of the H3 binding site could provide useful information to aid the development of allosteric small molecule modulators of KDM5A.


Carrier frequency analysis of mutations causing autosomal-recessive-inherited retinal diseases in the Israeli population.
Inherited retinal diseases (IRDs) are heterogeneous phenotypes caused by variants in a large number of genes. Disease prevalence and the frequency of carriers in the general population have been estimated in only a few studies, but are largely unknown. To this end, we developed two parallel methods to calculate carrier frequency for mutations causing autosomal-recessive (AR) IRDs in the Israeli population. We created an SQL database containing information on 178 genes from gnomAD (including genotyping of 5706 Ashkenazi Jewish (AJ) individuals) and our cohort of >2000 families with IRDs. Carrier frequency for IRD variants and genes was calculated based on allele frequency values and the Hardy-Weinberg (HW) equation. We identified 399 IRD-causing variants in 111 genes in Israeli patients and AJ controls. For the AJ subpopulation, gnomAD and HW-based regression analysis showed high correlation, therefore allowing one to use HW-based data as a reliable estimate of carrier frequency. Overall, carrier frequency per subpopulation ranges from 1/2.2 to 1/9.6 individuals, with the highest value obtained for the Arab-Muslim subpopulation in Jerusalem reaching an extremely high carrier rate of 44.7%. Carrier frequency per gene ranges from 1/31 to 1/11994 individuals. We estimate the total carrier frequency for AR-IRD mutations in the Israeli population as over 30%, a relatively high carrier frequency with marked variability among subpopulations. Therefore, these data are highly important for more reliable genetic counseling and genetic screening. Our method can be adapted to study other populations, either based on allele frequency data or cohort of patients.


Domain-specificity of Cd2+ and Zn2+ binding to rabbit liver metallothionein 2. Metal ion mobility in the formation of Cd4-metallothionein alpha-fragment.
The yield of the alpha-fragment of rabbit liver metallothionein 2 was used to test the domain-specificity and mobility of Cd2+ and Zn2+ when bound to metallothionein. Increasing molar ratios of Cd2+ were added to either Zn7-metallothionein or the metal-ion-free apo-metallothionein. The enzyme subtilisin was used to digest those parts of the peptide chain that were not bound to Cd2+. Analysis of the digestion products was carried out by separation by polyacrylamide-gel electrophoresis. The chelation agent EDTA was used as a competitive chelator. It was found that the presence of excess EDTA greatly enhances the formation of the Cd4-metallothionein alpha-fragment, and catalyses the complete digestion of all other the metal-ion-containing peptides, so that even Cd7-metallothionein, formed when 7 molar equivalents of Cd2+ are added to Zn7-metallothionein, is digested to the alpha-fragment. These results suggest that the Cd2+ bound in the beta-sites is very labile, much more labile than the kinetics of the off-reaction would suggest. The observation of significant amounts of alpha-fragment on the gels, even when the stoichiometry of the metal ions initially present in the protein should not have resulted in much concentration of Cd4-alpha-fragment clusters, indicates that as the digestion proceeds the metal ions move to sites that form complete clusters and therefore selectively protect that part of the peptide chain from digestion. We also find that rabbit Cd4-metallothionein 2 alpha-fragment stains near to the top of the gel, in complete contrast with the location of rat Cd4-metallothionein 2 alpha-fragment. This difference in the mobilities suggests that the alpha-fragment prepared from rabbit metallothionein 2 is much less negatively charged than the analogous protein fragment prepared from rat liver metallothionein 2.


What is the role of dose-dense therapy?
The introduction of paclitaxel/platinum combination chemotherapy and (interval) debulking surgery has significantly improved the prognosis of patients with ovarian cancer. Yet, many patients die of drug-resistant disease. Second-line chemotherapy may result in prolonged secondary remissions with alleviation of symptoms and improvement of quality of life. The response to second-line chemotherapy is strongly related to platinum sensitivity. More than 60% of platinum-sensitive patients respond to a re-challenge with platinum-containing chemotherapy. In platinum-resistant patients, on the contrary, the response rate to a re-challenge with 3-weekly platinum or any nonplatinum chemotherapy is less than 20%. The response to dose-dense weekly platinum-based regimens ranged from 48% to 64% in platinum-resistant patients. Moreover, the majority of the patients responded within 8 weeks after the start of the treatment. The progression-free survival ranged from a median of 5 months in a study using cisplatin/etoposide, to 11 months in a study with paclitaxel/carboplatin. The median survival was 11-15 months. The outpatient weekly paclitaxel/carboplatin regimen, with paclitaxel at a dose of 90 mg/m(2) and carboplatin at area under the curve 4, seems similarly effective and is better tolerated. Dose-dense weekly paclitaxel/carboplatin is an effective and well-tolerated therapy for platinum-sensitive, as well as platinum-resistant tumors. Responses to therapy are observed within 8 weeks in the majority of the patients. Whether a weekly regimen indeed is more effective than 3-weekly paclitaxel/carboplatin needs to be answered in a randomized study.


Clinical pharmacology of estrogens: cardiovascular actions and cardioprotective benefits of replacement therapy in postmenopausal women.
The use of estrogens in postmenopausal women has been the subject of much controversy regarding hormone formulation, dosage, use in combination with progestins, duration of treatment, and contraindications. Estrogens have been prescribed to relieve menopausal symptoms for more than three decades. The hormones reduce the gynecologic and psychologic changes associated with menopause while inhibiting bone resorption and possibly reducing the risk of cardiovascular disease. Their use however has been complicated by an increased risk of endometrial cancer and possibly breast cancer. The use of estrogens as cardioprotective agents is discussed and the clinical experiences and the possible mechanisms of action are reviewed. The clinical pharmacology of estrogens and the various formulations that are available as monotherapy or in combination with progestins will also be reviewed.


Compromised autophagy by MIR30B benefits the intracellular survival of Helicobacter pylori.
Helicobacter pylori evade immune responses and achieve persistent colonization in the stomach. However, the mechanism by which H. pylori infections persist is not clear. In this study, we showed that MIR30B is upregulated during H. pylori infection of an AGS cell line and human gastric tissues. Upregulation of MIR30B benefited bacterial replication by compromising the process of autophagy during the H. pylori infection. As a potential mechanistic explanation for this observation, we demonstrate that MIR30B directly targets ATG12 and BECN1, which are important proteins involved in autophagy. These results suggest that compromise of autophagy by MIR30B allows intracellular H. pylori to evade autophagic clearance, thereby contributing to the persistence of H. pylori infections.


A sensitive signal-on electrochemical assay for MTase activity using AuNPs amplification.
A sensitive and simple signal-on electrochemical assay for detection of Dam methyltransferase (MTase) activity based on DNA-functionalized gold nanoparticles (AuNPs) amplification coupled with enzyme-linkage reactions is presented. This new assay takes advantage of the steric hindrance of AuNPs and the electrostatic repulsion between the negative-charge phosphate backbones of DNA modified on the AuNPs and redox probe [Fe(CN)(6)](3-/4-). In this method, the self-assembled ssDNA on the electrode is hybridized with its complement ssDNA modified on AuNPs to form dsDNA AuNPs bioconjugates containing specific recognition sequence of Dam MTase and methylation-sensitive restriction endonuclease Dpn I. Then, the AuNPs approach to the electrode and result in blockage of electronic transmission. It is eT OFF state. In the presence of Dam MTase and Dpn I, the specific sequence is methylated and cleavaged, which in turn release the DNA modified AuNPs from the electrode surface allowing free exchange of electrons. It generates a measurable electrochemical signal (eT ON). Differential pulse voltammetry (DPV) is employed to detect the recover current, which is related to the concentration of the Dam MTase. This method is simple, sensitive, nonradioactive and without use of gel-electrophoresis, PCR or chromatographic separation. Under optimized conditions, a linear response to concentration of Dam MTase range from 0.2U/mL to 10 U/mL and a detection limit of 0.12 U/mL are obtained. Furthermore, our new assay is a promising method to detect Dam MTase in the Luria-Bertani (LB) medium, as well as to screen inhibitors or drugs for Dam MTase.


PATE, a gene expressed in prostate cancer, normal prostate, and testis, identified by a functional genomic approach.
To identify target antigens for prostate cancer therapy, we have combined computer-based screening of the human expressed sequence tag database and experimental expression analysis to identify genes that are expressed in normal prostate and prostate cancer but not in essential human tissues. Using this approach, we identified a gene that is expressed specifically in prostate cancer, normal prostate, and testis. The gene has a 1.5-kb transcript that encodes a protein of 14 kDa. We named this gene PATE (expressed in prostate and testis). In situ hybridization shows that PATE mRNA is expressed in the epithelial cells of prostate cancers and in normal prostate. Transfection of the PATE cDNA with a Myc epitope tag into NIH 3T3 cells and subsequent cell fractionation analysis shows that the PATE protein is localized in the membrane fraction of the cell. Analysis of the amino acid sequence of PATE shows that it has structural similarities to a group of proteins known as three-finger toxins, which includes the extracellular domain of the type beta transforming growth factor receptor. Restricted expression of PATE makes it a potential candidate for the immunotherapy of prostate cancer.


Acute lymphoblastic leukaemia in a child with systemic lupus erythematosus.
Haematological involvement of systemic lupus erythematosus (SLE) - which ranges from the well-described haemolytic anaemia to macrophage activation syndrome - has a large impact on both morbidity and mortality. On the other hand, association between haematological malignities and SLE - in terms of pathophysiology and molecular genetics - is an obscure entity which has not been clarified evidently to date. Herein, we present a six-year-old female with the diagnosis of SLE who developed acute lymphoblastic leukaemia following a period of myelodysplasia. It could possibly be coincidental; however, persistent cytopenia, prominent dysplasia on bone marrow smears and azathioprine treatment may be considered as possible triggers for the development of leukaemia in the present case.


Prevalence of clinical signs of possible serious bacterial infection and mortality associated with them from population-based surveillance of young infants from birth to 2 months of age.
Community-based data on the prevalence of clinical signs of possible serious bacterial infection (PSBI) and the mortality associated with them are scarce. The aim was to examine the prevalence for each sign of infection and mortality associated with infants in the first two months of life, using community surveillance through community health workers (CHW). We used population-based surveillance data of infants up to two months of age from the African Neonatal Sepsis Trial (AFRINEST). In this study, CHWs visited infants up to 10 times during the first two months of life at five sites in three sub-Saharan African countries. CHW assessed the infant for signs of infection (local or systemic) and referred infants who presented with any sign of infection to a health facility. We used a longitudinal analysis to calculate the risk of death associated with the presence of a sign of infection at the time of the visit until the subsequent visit. During the first two months of their life, CHWs visited 84,759 live-born infants at least twice. In 11,089 infants (13.1%), one or more signs of infection were identified, of which 237 (2.1%) died. A sign of infection was detected at 2.1% of total visits. In 52% of visits, infants had one or more sign of systemic infection, while 25% had fast breathing in 7-59 days period and 23% had a local infection. All signs of infection, including multiple signs, were more frequently seen in the first week of life. The risk of mortality was very low (0.2%) for local infections and fast breathing in 7-59 days old, it was low for fast breathing 0-6 days old (0.6%), high body temperature (0.7%) and severe chest indrawing (1.0%), moderate for low body temperature (4.9%) and stopped feeding well/not able to feed at all (5.0%) and high for movement only when stimulated or no movement at all (10%) and multiple signs of systemic infection (15.5%). The risk of death associated with most clinical signs was higher (1.5 to 9 times) in the first week of life than at later age, except for low body temperature (4 times lower) as well as high body temperature (2 times lower). Signs of infections are common in the first two months of life. The mortality risk differs with clinical signs and can be grouped as very low (local infections, fast breathing 7-59 days), low (fever, severe chest indrawing and fast breathing 0-6 days), moderate (low body temperature and stopped feeding well/not able to feed at all) and high (for movements only on stimulation or no movements at all and multiple signs of infection). New treatment strategies that consider differential mortality risk could be developed and evaluated based on these findings. The trial was registered with Australian New Zealand Clinical Trials Registry under ID ACTRN 12610000286044.


Predictive factors for missed adenoma on repeat colonoscopy in patients with suboptimal bowel preparation on initial colonoscopy: A KASID multicenter study.
Suboptimal bowel preparation can result in missed colorectal adenoma that can evolve into interval colorectal cancer. This study aims to identify the predictive factors associated with missed adenoma on repeat colonoscopy in patients with suboptimal bowel preparation at initial colonoscopy. A total of 441 patients with suboptimal bowel preparation on initial colonoscopy and who had repeat colonoscopy within two years were included from 2007 to 2014 in six tertiary hospitals. Suboptimal bowel preparation was defined as 'poor' according to the Aronchick scale or a score ≤ 1 in at least one segment or total score < 6 according to the Boston bowel preparation scale. Of 441 patients, mean age at initial colonoscopy was 59.1 years, and 69.2% patients were male. The mean interval from initial to repeat colonoscopy was 14.1 months. The per-patient adenoma miss rate (AMR) was 42.4% for any adenoma and 5.4% for advanced adenoma. When the association between baseline clinical characteristics and missed lesions on repeat colonoscopy was analyzed, dyslipidemia (odds ratio [OR], 5.19; 95% confidence interval [CI], 1.14-23.66; P = 0.034), and high-risk adenoma (OR, 4.45; 95% CI, 1.12-17.68; P = 0.034) on initial colonoscopy were independent risk factors for missed advanced adenoma. In patients with suboptimal bowel preparation, dyslipidemia and high-risk adenoma on initial colonoscopy were independently predictive of missed advanced adenoma on repeat colonoscopy.


DNA protein interactions at the rRNA of Saccharomyces cerevisiae.
The rDNA cluster is the genetic locus encoding the ribosomal RNAs and physically defines where ribosomes begin to be assembled. In the yeast Saccharomyces cerevisiae, the highly repetitive structure of this locus makes it a very interesting target for studies about genome stability, chromatin-mediated transcriptional silencing and progression of aging. In fact, recombination among the repeated units is suppressed in a WT cell. Moreover, when genes transcribed by RNA polymerase II are inserted in the rDNA cluster, their transcription is silenced. Finally, the formation of rDNA minicircles (ERCs) has been shown to be one of the causes of aging in yeast. DNA topoisomerase I have been shown to suppress recombination specifically at the rDNA of S.cerevisiae. Moreover, also the chromatin structure of this locus is affected in a top1 strain, because rDNA specific transcriptional silencing is abolished. Nonetheless, the molecular basis of how this enzyme interferes with these functions is yet unknown. Here are reported results obtained by in vivo studies of DNA protein interactions occurring on the rDNA locus. The analyses include a fine mapping of nucleosome positioning; RNA polymerase I transcription factors and DNA topoisomerase I cleavage sites. Important conclusions can be drawn: i) nucleosome positioning in the Non Transcribed Spacer is not affected by RNA polymerase I transcription; ii) the RNA polymerase I transcription factors bind DNA in vivo with a defined hierarchy; iii) the DNA topoisomerase I cleaves the NTS in very specific sites, but cleavage is not induced by RNA polymerase I transcription. These in vivo studies help to characterize the molecular basis of important phenomena as the transcriptional silencing and genome stability in yeast.


A Pilot, Virtual Exercise Intervention Improves Health and Fitness during the COVID-19 Pandemic.
Physical activity levels are low in individuals with chronic disease (e.g., obesity) and have worsened during the COVID-19 pandemic. Our pilot study tested a virtual exercise intervention for rural-dwelling adults with chronic disease from January-April 2021 for changes in mental health, physical fitness, and physical activity and for intervention fidelity. Participants (n = 8 [7 female]; age = 57.5 ± 13.8 years, body mass index = 38.2 ± 8.0 kg/m2) completed an exercise intervention led virtually by collegiate health science majors. Participants attended two 60-minute sessions/week for 12 weeks, completing individually-tailored and progressed aerobic and muscle-strengthening training. A non-randomized control group matched on gender and age continued normal activity during the 12 weeks. Changes in mental health, physical fitness, and physical activity measures were evaluated using a 2×2 (group × time) analysis of covariance. Both groups improved mental health, but only intervention participants lost weight (3.1 ± 1.0 kg; no change in controls). Step test, arm curls, and chair stands improved by 16.1-20.6% in the intervention and 7.8-12.1% in the control groups. Intervention participants did not increase overall physical activity during or after the intervention. Intervention fidelity was high; participants attended ~73% of sessions and rated the sessions 4.7 ± 0.6 (out of 5). Researcher observations rated exercise sessions as meeting 12.7 ± 0.6 of 16 goals. Our virtual exercise program was associated with positive mental health and physical fitness changes. Such programs may provide a method, even beyond the pandemic, to improve fitness in adults with chronic disease.


[Genetic counseling of acrocephalosyndactylia].
The present paper describes a case with clinical and radiological characteristics of Apert's syndrome. He was the first offspring from second cousins marriage. We considered that in this case, consanguinity is an associated feature. Perhaps this case may be the third one reported in the literature with consanguineous parents. We comment the different classifications of the acrocephalosyndactylias for genetic counseling purposes.


Clinicopathological and prognostic features of operable non-small cell lung cancer patients with diabetes mellitus.
The aim of this study was to investigate the clinicopathological and prognostic features of operable non-small cell lung cancer (NSCLC) patients with diabetes mellitus (DM). A total of 1231 surgically resected NSCLC patients were retrospectively reviewed. Clinicopathological characteristics were compared between patients with DM (DM group, n = 139) and those without DM (non-DM group, n = 1092). The clinical factors associated with postoperative complications and prognostic factors were identified. The DM group had distinct clinicopathological features. No significant differences in histological invasiveness or stage were found. The presence and control status of DM were independent predictors of postoperative complications. No significant differences in recurrence-free survival or cancer-specific survival were observed; however, the DM group had worse overall survival (OS). The DM group had a higher number of deaths from other diseases than the non-DM group, and these patients had significantly higher postoperative hemoglobin A1c levels than patients with cancer-related death. The presence and control status of preoperative DM are useful predictors of both postoperative complications and OS in operable NSCLC patients. Concomitant diabetes-related complications have a negative effect on long-term survival in diabetic NSCLC patients, and long-term glycemic control is important to prolong OS.


Craniofacial development: discoveries made in the chicken embryo.
The aim of this review is to highlight some of the key contributions to our understanding of craniofacial research from work carried out with the chicken and other avian embryos. From the very first observations of neural crest cell migration to the fusion of the primary palate, the chicken has proven indispensable in facilitating craniofacial research. In this review we will look back to the premolecular studies where "cut and paste" grafting experiments mapped the fate of cranial neural crest cells, the role of different tissue layers in patterning the face, and more recently the contribution of neural crest cells to jaw size and identity. In the late 80's the focus shifted to the molecular underpinnings of facial development and, in addition to grafting experiments, various chemicals and growth factors were being applied to the face. The chicken is above all else an experimental model, inviting hands-on manipulations. We describe the elegant discoveries made by directly controlling signaling either in the brain, in the pharyngeal arches or in the face itself. We cover how sonic hedgehog (Shh) signals to the face and how various growth factors regulate facial prominence identity, growth and fusion. We also review abnormal craniofacial development and how several type of spontaneous chicken mutants shed new light on diseases affecting the primary cilium in humans. Finally, we bring out the very important role that the bird beak has played in understanding amniote evolution. The chicken, duck and quail have been and will continue to be used as experimental models to explore the evolution of jaw diversity and the morphological constraints of the vertebrate face.


REV-ERBα Regulates CYP7A1 Through Repression of Liver Receptor Homolog-1.
Nuclear heme receptor reverse erythroblastosis virus (REV-ERB) α (a transcriptional repressor) is known to regulate cholesterol 7α-hydroxylase (CYP7A1) and bile acid synthesis. However, the mechanism for REV-ERBα regulation of CYP7A1 remains elusive. Here, we investigate the role of LRH-1 in REV-ERBα regulation of CYP7A1 and cholesterol metabolism. We first characterized the tertiary amine N-(4-chloro-2-methylbenzyl)-N-(4-chlorobenzyl)-1-(5-nitrothiophen-2-yl)methanamine (GSK2945) as a highly specific Rev-erbα/REV-ERBα antagonist using cell-based assays and confirmed expression of Rev-erbα in mouse liver. GSK2945 treatment increased hepatic mouse cholesterol 7α-hydroxylase (Cyp7a1) level and lowered plasma cholesterol in wild-type mice. Likewise, the compound increased the expression and microsomal activity of Cyp7a1 in hypercholesterolemic mice. This coincided with reduced plasma and liver cholesterol and enhanced production of bile acids. Increased levels of Cyp7a1/CYP7A1 were also found in mouse and human primary hepatocytes after GSK2945 treatment. In these experiments, we observed parallel increases in Lrh-1/LRH-1 (a known hepatic activator of Cyp7a1/CYP7A1) mRNA and protein. Luciferase reporter, mobility shift, and chromatin immunoprecipitation assays revealed that Lrh-1/LRH-1 was a direct Rev-erbα/REV-ERBα target gene. Furthermore, conditional deletion of Lrh-1 in the liver abrogated the regulatory effects of Rev-erbα on Cyp7a1 and cholesterol metabolism in mice. In conclusion, Rev-erbα regulates Cyp7a1 and cholesterol metabolism through its repression of the Lrh-1 receptor. Targeting the REV-ERBα/LRH-1 axis may represent a novel approach for management of cholesterol-related diseases.


The influence of pH on membrane conductance and intercellular resistance in the rat lens.
1. The conductance of the rat lens was measured using a two-internal-microelectrode technique. The voltage response to a step of current consisted of two components arising from bulk and membrane resistance respectively. 2. The potassium permeability was calculated by applying Goldman theory to 86Rb+ efflux data. 3. The internal pH (pHi) and internal free calcium (pCai) were measured directly using single- and double-barrelled ion-sensitive microelectrodes. 4. Lens pHi was 6.9 in control solution (external pH, pHo = 7.3) and was reduced on lowering pHo. The presence of propionate or 100% CO2 in the external solution accentuated this effect. 5. Internal acidification was accompanied by a depolarization of membrane potential, an increase in membrane and cell-to-cell resistance and a decrease in potassium permeability. The acidification had no effect on pCai. 6. The intracellular pH was increased by perifusing with trimethylamine or NH4Cl. Both treatments induced a membrane depolarization with little change in potassium permeability. Subsequent removal of NH4Cl led to a sustained decrease in pHi. 7. In every case where pHi decreased, the changes in membrane potential and conductance could be explained largely on the basis of a decrease in potassium permeability. The concomitant increase in cell-to-cell resistance was less pronounced and probably insufficient to uncouple the lens system.


Postoperative control for the persistence of positivity in three immunological tests used in the diagnosis of hydatidosis in children (E.L.I.S.A.-specific IgE, indirect hemagglutination and E.L.I.S.A.-total IgE).
The persistence of serological positivity was studied in 21 children with surgically proven hydatidosis. The children had been previously operated on between 10 months and 7 4/12 years before serum samples were extracted. Forty-eight people (19 adult and 39 children) were included as a control group. E.L.I.S.A. specific IgE and indirect hemagglutination techniques were used for the diagnosis of hydatid disease. Studies of the immunoglobulins and complement components IgG, IgA, IgM, IgE, C3, C4, C5 and factor B were done on the sera from the 21 children with histories of equinococciasis. The results are expressed according to the location and physical condition of the cyst, the time passed after surgery, the number of cysts and the number of surgical operations. We found persistence of positivity of up to 5 4/12 years for indirect hemagglutination and up to 6 7/12 years for specific IgE, in cases without evidence of clinical parasitation so we think that the postoperative control must be based on several serological determinations. The concordance between the two specific serological tests (specific IgE and indirect hemagglutination) was 80.95%; specific IgE having appeared more sensitive especially in cases of hepatic location. The results concerning the total IgE determination were highlighted in the immunoglobulin studies. We consider total IgE as being only orientative, because of the frequency with which it is found elevated in childhood diseases.


Anatomical basis and physiological role of cerebrospinal fluid transport through the murine cribriform plate.
Cerebrospinal fluid (CSF) flows through the brain, transporting chemical signals and removing waste. CSF production in the brain is balanced by a constant outflow of CSF, the anatomical basis of which is poorly understood. Here, we characterized the anatomy and physiological function of the CSF outflow pathway along the olfactory sensory nerves through the cribriform plate, and into the nasal epithelia. Chemical ablation of olfactory sensory nerves greatly reduced outflow of CSF through the cribriform plate. The reduction in CSF outflow did not cause an increase in intracranial pressure (ICP), consistent with an alteration in the pattern of CSF drainage or production. Our results suggest that damage to olfactory sensory neurons (such as from air pollution) could contribute to altered CSF turnover and flow, providing a potential mechanism for neurological diseases.


Discriminatory power of tests applied in back pain during pregnancy.
A longitudinal, prospective, observational cohort study. To assess the relationship between clinical back status and reported pain locations during and after pregnancy. Back pain during pregnancy is a frequent clinical occurrence, even during the early stages of pregnancy. The cause is unclear. There are few data describing the results of a general physical examination of the back during pregnancy and there are no data on serial examinations. Such data could provide information about what structures cause the pain, which might have implications for the choice of treatment. A cohort of 200 consecutive women attending an antenatal clinic was observed throughout the pregnancy terms, and repeated measurements of back pain and its possible determinants were taken using questionnaires and physical examinations in a standardized way, including a series of tests of configuration, mobility, and pain provocation. Pain provocation tests were better at discriminating among women who reported back pain from women who reported no back pain from tests of configuration or mobility. The discriminatory power of the tests was better in the lower part of the spine than in the upper part. The best discrimination was achieved by combining some of the tests. The results indicate that not one but several pain-releasing structures may be involved. These are probably the various pelvic ligaments, which may form a functional unit. These findings may have therapeutic implications.


Rab28 function in trypanosomes: interactions with retromer and ESCRT pathways.
Early endosomal cargo is typically targeted to either a degradative or recycling pathway. Despite established functions for the retromer and ESCRT complexes at late endosomes/multivesicular bodies, the mechanisms integrating and coordinating these functions remain largely unknown. Rab family GTPases are key membrane trafficking organizers and could contribute. Here, in the unicellular organism Trypanosoma brucei, we demonstrate that Rab28 locates to the endosomal pathway and partially colocalizes with Vps23, an ESCRT I component. Rab28 is required for turnover of endocytosed proteins and for lysosomal delivery of protein cargo. Using RNA interference we find that in Rab28-depleted cells, protein levels of ESCRT I (Vps23/28) and retromer (Vps26) are also decreased, suggesting that Rab28 is an important regulator of these factors. We suggest that Rab28 coordinates the activity of retromer-dependent trafficking and ESCRT-mediated degradative pathways.


Automatic breach detection during spine pedicle drilling based on vibroacoustic sensing.
Pedicle drilling is a complex and critical spinal surgery task. Detecting breach or penetration of the surgical tool to the cortical wall during pilot-hole drilling is essential to avoid damage to vital anatomical structures adjacent to the pedicle, such as the spinal cord, blood vessels, and nerves. Currently, the guidance of pedicle drilling is done using image-guided methods that are radiation intensive and limited to the preoperative information. This work proposes a new radiation-free breach detection algorithm leveraging a non-visual sensor setup in combination with deep learning approach. Multiple vibroacoustic sensors, such as a contact microphone, a free-field microphone, a tri-axial accelerometer, a uni-axial accelerometer, and an optical tracking system were integrated into the setup. Data were collected on four cadaveric human spines, ranging from L5 to T10. An experienced spine surgeon drilled the pedicles relying on optical navigation. A new automatic labeling method based on the tracking data was introduced. Labeled data was subsequently fed to the network in mel-spectrograms, classifying the data into breach and non-breach. Different sensor types, sensor positioning, and their combinations were evaluated. The best results in breach recall for individual sensors could be achieved using contact microphones attached to the dorsal skin (85.8%) and uni-axial accelerometers clamped to the spinous process of the drilled vertebra (81.0%). The best-performing data fusion model combined the latter two sensors with a breach recall of 98%. The proposed method shows the great potential of non-visual sensor fusion for avoiding screw misplacement and accidental bone breaches during pedicle drilling and could be extended to further surgical applications.


Quantitative tract-based white matter development from birth to age 2years.
Few large-scale studies have been done to characterize the normal human brain white matter growth in the first years of life. We investigated white matter maturation patterns in major fiber pathways in a large cohort of healthy young children from birth to age two using diffusion parameters fractional anisotropy (FA), radial diffusivity (RD) and axial diffusivity (RD). Ten fiber pathways, including commissural, association and projection tracts, were examined with tract-based analysis, providing more detailed and continuous spatial developmental patterns compared to conventional ROI based methods. All DTI data sets were transformed to a population specific atlas with a group-wise longitudinal large deformation diffeomorphic registration approach. Diffusion measurements were analyzed along the major fiber tracts obtained in the atlas space. All fiber bundles show increasing FA values and decreasing radial and axial diffusivities during development in the first 2years of life. The changing rates of the diffusion indices are faster in the first year than the second year for all tracts. RD and FA show larger percentage changes in the first and second years than AD. The gender effects on the diffusion measures are small. Along different spatial locations of fiber tracts, maturation does not always follow the same speed. Temporal and spatial diffusion changes near cortical regions are in general smaller than changes in central regions. Overall developmental patterns revealed in our study confirm the general rules of white matter maturation. This work shows a promising framework to study and analyze white matter maturation in a tract-based fashion. Compared to most previous studies that are ROI-based, our approach has the potential to discover localized development patterns associated with fiber tracts of interest.


Verteporfin-loaded microparticles for radiosensitization of preclinical lung and breast metastatic spine cancer.
The vertebral column is the most common site for skeletal metastasis, often leading to debilitating pain and weakness. Metastatic cancer has unique genetic drivers that potentiate tumorigenicity. There is an unmet need for novel targeted therapy in patients with spinal metastatic disease. The authors assessed the effect of verteporfin-induced yes-associated protein (YAP) inhibition on spine metastatic cell tumorigenicity and radiation sensitivity in vitro. Animal studies used a subcutaneous xenograft mouse model to assess the use of systemic intraperitoneal verteporfin (IP-VP) and intratumoral verteporfin microparticles (IT-VP) to inhibit the tumorigenicity of lung and breast spinal metastatic tumors from primary patient-derived tissue. Verteporfin led to a dose-dependent decrease in migration, clonogenicity, and cell viability via inhibition of YAP and downstream effectors cyclin D1, CTGF, TOP2A, ANDRD1, MCL-1, FOSL2, KIF14, and KIF23. This was confirmed with knockdown of YAP. Verteporfin has an additive response when combined with radiation, and knockdown of YAP rendered cells more sensitive to radiation. The addition of verteporfin to YAP knockdown cells did not significantly alter migration, clonogenicity, or cell viability. IP-VP and IT-VP led to diminished tumor growth (p < 0.0001), especially when combined with radiation (p < 0.0001). Tissue analysis revealed diminished expression of YAP (p < 0.0001), MCL-1 (p < 0.0001), and Ki-67 (p < 0.0001) in tissue from verteporfin-treated tumors compared with vehicle-treated tumors. This is the first study to demonstrate that verteporfin-mediated inhibition of YAP leads to diminished tumorigenicity in lung and breast spinal metastatic cancer cells. Targeting of YAP with verteporfin offers promising results that could be translated to human clinical trials.


A variety of gene polymorphisms associated with idiopathic granulomatous mastitis.
Idiopathic granulomatous mastitis (IGM) is a rare and chronic inflammatory disorder. IGM mimics breast cancer regarding its clinical and radiological features. Etiology of IGM remains unclarified. Our patient was 37-year-old and 14 weeks pregnant. There was pain, redness and swelling in the right breast. The mass suggestive of malignancy was detected in sonography. Serum CA 125 and CA 15-3 levels were high. Genetic analysis was performed for the etiology. methylenetetrahydrofolate reductase (MTHFR) C 677 TT, β-fibrinogen-455 G>A, plasminogen activator inhibitor (PAI)-1 5 G/5 G, angiotensin-converting enzyme (ACE) I/D mutation was found. IGM was diagnosed by cor biopsy. An association was also reported between breast cancer and mutations in MTHFR-C 677 T, PAI-1, ACE genes. Genetic polymorphisms may involve in the development of IGM as it was seen in our case. Further studies should be conducted to better clarify this plausible association.


Health services use among formerly incarcerated Louisiana Medicaid members within one year of release.
To determine the association between enrollment in Medicaid prior to release compared with post-release, and the use of health services and time to the first service use after release among Louisiana Medicaid members within one year of release from Louisiana state corrections custody. We conducted a retrospective cohort study linking Louisiana Medicaid and Louisiana state corrections release data. We included individuals ages 19 to 64 years released from state custody between January 1, 2017 and June 30, 2019 and enrolled in Medicaid within 180 days of release. Outcome measures included receipt of general health services (primary care visits, emergency department visits, and hospitalizations), cancer screenings, specialty behavioral health services, and prescription medications. To determine the association between pre-release Medicaid enrollment and time to receipt of health services, multivariable regression models were used which accounted for significant differences in characteristics between the groups. Overall, 13283 individuals met eligibility criteria and 78.8% (n = 10473) of the population was enrolled in Medicaid pre-release. Compared with those enrolled in Medicaid prior to release, those enrolled post-release were more likely to have an emergency department visit (59.6% versus 57.5%, p = 0.04) and hospitalization (17.9% versus 15.9%, p = 0.01) and less likely to receive outpatient mental health services (12.3% versus 15.2%, p<0.001) and prescription drugs. Compared with those enrolled in Medicaid prior to release, those enrolled post-release had a significantly longer time to receiving many services including a primary care visit (adjusted mean difference: 42.2 days [95% CI: 37.9 to 46.5; p<0.001]), outpatient mental health services (42.8 days [95% CI: 31.3 to 54.4; p<0.001]), outpatient substance use disorder service (20.6 days [95% CI: 2.0 to 39.2; p = 0.03]), and medication for opioid use disorder (40.4 days [95% CI: 23.7 to 57.1; p<0.001]) as well as inhaled bronchodilators and corticosteroids (63.8 days [95% CI: 49.3 to 78.3, p<0.001]), antipsychotics (62.9 days [95% CI: 50.8 to 75.1; p<0.001]), antihypertensives (60.5 days [95% CI: 50.7 to 70.3; p<0.001]), and antidepressants (52.3 days [95% CI: 44.1 to 60.5; p<0.001]). Compared with Medicaid enrollment post-release, pre-release Medicaid enrollment was associated with higher proportions of, and faster access to, a wide variety of health services. Regardless of enrollment status, we found prolonged times between release and receipt of time-sensitive behavioral health services and prescription medications.


Altered anterograde axonal transport of mitochondria in cultured striatal neurons of a knock-in mouse model of Huntington's disease.
Huntington's disease (HD) is a progressive genetic neurodegenerative disease caused by an abnormal expansion of a cytosine-adenine-guanine trinucleotide repeat in the huntingtin gene. One pathological feature of HD is neuronal loss in the striatum. Despite many efforts, mechanisms underlying neuronal loss in HD striatum remain elusive. It was suggested that the mutant huntingtin protein interacts mitochondrial proteins and causes mitochondrial dysfunction in striatal neurons. However, whether axonal transport of mitochondria is altered in HD striatal neurons remains controversial. Here, we examined axonal transport of single mitochondria labelled with Mito-DsRed2 in cultured striatal neurons of zQ175 knock-in mice (a knock-in mouse model of HD). We observed decreased anterograde axonal transport of proximal mitochondria in HD striatal neurons compared with wild-type (WT) striatal neurons. Decreased anterograde transport in HD striatal neurons was prevented by overexpressing mitochondrial Rho GTPase 1 (Miro1). Our results offer a new insight into mechanisms underlying neuronal loss in the striatum in HD.


Complementary and alternative medicine (CAM) in Mexican patients with cancer.
Cancer is the second cause of death in Mexico, most cases are detected at advanced stages and the use of chemotherapy is frequent. At present, more than 300 types of complementary and/or alternative medicine (CAM) treatments are known that offer different therapeutic objectives. Many patients use this type of treatments. To determine the characteristics of the patients that use CAM, to identify the aim of these treatments, the source of information and the potential benefits obtained by the patient. A questionnaire was applied from February 20 to March 5, 2004 to non-selected patients with cancer in private consultation to determine age, sex, education level, work, use of CAM, type and number of used therapies, potential benefits and monthly cost. Two groups were formed, A for users and B for non-users of CAM. Results between groups were compared and the mentioned variables were correlated with the use of alternative medicine. Group A included 37 patients and group B included 38, with no difference regarding age, education level, work and oncological diagnosis, p > 0.05; a significant trend was found as regards the feminine sex, p = 0.07, neoplasm different from breast cancer, p = 0.08 and evident association with neoplasm advanced stages, p = 0.02. Most patients used between 1 and 3 types of therapies, 97.2%. The most common types of therapies were nutritional and spiritual, 54% and 48.6%, respectively. The source was the patient's family in 56.4% and the physician in 24.3%. Complementary and alternative therapy was considered a success in 57.1%; most of the users mentioned benefits (78.6%) deemed as tranquility (46.4%) or improvement of the physical condition (46.4%). The average monthly cost was $ 345.5 dollars, with a range of $ 13.6 to $ 2,545.5 dollars. The use of complementary and/or alternative therapy is frequent among young women with advanced cancer and high level of education. The family participates in the decision of using these methods; most users noticed a benefit in their general condition and reported tranquility; these patients may be prone to higher incidence of depression and anxiety. The effectiveness and safety of this type of treatments remain to be determined, as well as the possible interactions with conventional therapy.


The effects of age and delay on responses to repeated questions in forensic interviews with children alleging sexual abuse.
We examined transcripts of forensic interviews with 115 children aged between 3 and 12 years, interviewed between 1 day and 18 months after allegedly experiencing a single incident of sexual abuse. Repeated questions were categorized with respect to the reasons why interviewers asked questions again, how interviewers asked repeated questions, and how children responded. On average, interviewers asked 3 repeated questions per interview. As age increased, the frequency of question repetition declined but there was no association between repetition and delay. Interviewers most often repeated questions for clarification (53.1%), but questions were also repeated frequently to challenge children's previous responses (23.7%), and for no apparent reason (20.1%). In response, children typically repeated (54.1%) or elaborated on (31.5%) their previous answers; they contradicted themselves less often (10.8%). Questions repeated using suggestive prompts were more likely to elicit contradictions. There was no association between age or delay and the reasons why questions were repeated, how they were repeated, and how children responded. These findings emphasize the importance of training forensic interviewers to repeat questions only when the children or interviewers seek clarification and to encourage children who are anxious or reluctant to disclose. All repeated questions should be open-ended and interviewers should explain to children why questions are being repeated.


Immunotoxicity of bisphenol S and F are similar to that of bisphenol A during zebrafish early development.
Bisphenol S (BPS) and bisphenol F (BPF) have been increasingly used as alternatives to bisphenol A (BPA) owing to health concerns. The present study aims to evaluate the impact of these two BPA analogs on oxidative stress and the immune system during zebrafish embryonic and larval development. Environmentally relevant levels of BPS and BPF exposure could increase reactive oxygen species (ROS) content, nitric oxide (NO) content, nitric oxide synthase (NOS) activity, and the expression of immunity-related genes in concentration dependent manners during the early developmental stages in zebrafish. At a concentration of 100 μg/L, BPS and BPF showed similar effects on the immune toxicity of zebrafish as that of BPA. Moreover, BPS and BPF induced both erα and nf-κb expression, and antagonists of estrogen receptor and NF-κB blocked the effects on immunity-related gene expression, providing evidence that the two pathways mediate the actions of BPS and BPF on fish immune response and functions. Thus we conclude that the presence of BPS and BPF in the environment, similar to BPA, may also pose risks to ecosystem and human health and cannot be widely used without limitations and precautions.


Time reflection and refraction of graphene plasmons at a temporal discontinuity.
A dynamical change of material properties induces a special type of reflection and refraction at a temporal discontinuity. Here, we study the interaction of graphene plasmons with single and double temporal discontinuities or shocks, leading to controlled in-plane scattering. We analytically determine the Fresnel-like coefficients for graphene plasmons at these boundaries, and validate our results by rigorous numerical simulations. Temporally controlled doping of two-dimensional materials such as graphene thus leads to a new mechanism for planar and compact plasmonic devices.


Emotion regulation strategies in mania risk: A systematic review.
Difficulties in emotion regulation may contribute to the development of mania. This review aimed to assess how emotion regulation strategies reported by individuals at risk of mania compare with clinical and nonclinical controls. Search terms relating to mania risk and emotion regulation were entered into three databases. Sixteen studies were included. Mania risk was typically associated with overall endorsement of emotion regulation strategies, particularly dampening, and positive and negative rumination. Findings were limited by overall lack of evidence for individual strategies, lack of consideration of key mediating factors and reliance upon self-report designs.


Whole-genome SNP association analysis of reproduction traits in the Finnish Landrace pig breed.
Good genetic progress for pig reproduction traits has been achieved using a quantitative genetics-based multi-trait BLUP evaluation system. At present, whole-genome single nucleotide polymorphisms (SNP) panels provide a new tool for pig selection. The purpose of this study was to identify SNP associated with reproduction traits in the Finnish Landrace pig breed using the Illumina PorcineSNP60 BeadChip. Association of each SNP with different traits was tested with a weighted linear model, using SNP genotype as a covariate and animal as a random variable. Deregressed estimated breeding values of the progeny tested boars were used as the dependent variable and weights were based on their reliabilities. Statistical significance of the associations was based on Bonferroni-corrected P-values. Deregressed estimated breeding values were available for 328 genotyped boars. Of the 62,163 SNP in the chip, 57,868 SNP had a call rate > 0.9 and 7,632 SNP were monomorphic. Statistically significant results (P-value < 2.0E-06) were obtained for total number of piglets born in first and later parities and piglet mortality between birth and weaning in later parity, and suggestive associations (P-value < 4.0E-06) for piglet mortality between birth and weaning in first parity, number of stillborn piglets in later parity, first farrowing interval and second farrowing interval. Two of the statistically significant regions for total number of piglets born in first and later parities are located on chromosome 9 around 95 and 79 Mb. The estimated SNP effect in these regions was approximately one piglet between the two homozygote classes. By combining the two most significant SNP in these regions, favourable double homozygote animals are expected to have 1.3 piglets (P-value = 1.69E-08) more than unfavourable double homozygote animals. A region on chromosome 9 (66 Mb) was statistically significant for piglet mortality between birth and weaning in later parity (0.44 piglets between homozygotes, P-value = 6.94E-08). Three separate regions on chromosome 9 gave significant results for litter size and pig mortality. The frequencies of favourable alleles of the significant SNP are moderate in the Finnish Landrace population and these SNP are thus valuable candidates for possible marker-assisted selection.


The ICLAS/CCAC International Symposium on Regulatory Testing and Animal Welfare.
The first International Symposium on Regulatory Testing and Animal Welfare (ISRTAW), held 21-23 June 2001, in Quebec City, Canada, brought together 160 experts from 22 countries from North and South America, Europe and Asia. The experts included representatives from national research and regulatory agencies, universities, and industry involved in chemicals, pesticides and drug safety testing. Representatives from European, Canadian and US animal welfare groups also participated in the discussions. The Symposium was organised by the International Council for Laboratory Animal Science (ICLAS) and the Canadian Council on Animal Care (CCAC), with the support and assistance of many sponsors and advisors. ICLAS is a worldwide organisation whose purpose is to foster the international harmonisation of animal care and use practices. CCAC is the national agency responsible for overseeing the ethical use of animals in Canadian science. Both organisations are committed to fostering an environment in which global efforts to harmonise testing procedures using animals in a more-humane manner can be realised.


Thiabendazole-induced acute liver failure requiring transplantation and subsequent diagnosis of polyarteritis nodosa.
Polyarteritis nodosa (PAN), a systemic necrotising vasculitis that affects medium- and small-sized arteries, has visceral involvement in 40-60% of the patients. According to the Five-Factor Score (FFS), it is associated with poor outcome. We describe a patient who underwent orthotopic liver transplantation (OLT) for severe ductopenia induced by thiabendazole that was empirically prescribed for chronic hypereosinophilia. Eleven years later, despite immunosuppressive treatment to prevent graft rejection, he developed mononeuritis multiplex; PAN was diagnosed. He also had severe recurrent ischaemic cholangitides because of post-OLT hepatic artery ligation to treat a postoperative severe haematemesis. His outcome was favourable after second OLT, under steroids, cyclophosphamide pulses and tacrolimus. In retrospect, his initial symptoms and hypereosinophilia were probably attributable to PAN.


A method for computing the partial singular value decomposition.
A method for computing the partial singular value decomposition of a matrix is described. The method is appropriate to problems where the matrix is known to be of low rank and only the principal singular vectors are of interest. The technique is simple, easy to implement in integer arithmetic, and places modest memory requirements. The convergence properties of the algorithm are investigated analytically and by simulation.


Molecular identification and expression of Lyn tyrosine kinase isoforms in marsupials.
Lyn is a tyrosine kinase molecule required for modulation of signalling cascades in cell populations including B lymphocytes of the mammalian immune system. We have characterised the coding domain of the marsupial lyn gene of two macropod marsupials; the Tammar Wallaby (Macropus eugenii) and the Bridled Nailtail Wallaby (Onychogalea fraenata) and show the co-expression of two Lyn isoforms in cells and tissues of these and three other marsupials (Brush-tail Possum, Trichosurus vulpecula; American Grey Short-tailed Opossum, Monodelphis domestica and Red-tailed Phascogale, Phascogale calura). The predicted Lyn proteins (LynA and LynB) were highly conserved across vertebrate species, with amino acid identities of 94% with their human orthologues and conservation of key tyrosine kinase motifs that suggests that marsupial Lyn most likely functions in cell signalling. Comparison of our cDNA data to annotations for Lyn transcripts (available through the Ensembl Genome Browser) for the Tammar Wallaby confirm splice sites for a number of exons in the wallaby transcript that are missing from the current annotation. This is the first report of the expression of kinase signalling molecules that influence immunity in metatherian mammals and provides key information to support ongoing studies of immune regulation in marsupials.


JmjC Domain-Encoding Genes Are Conserved in Highly Regenerative Metazoans and Are Associated with Planarian Whole-Body Regeneration.
The capacity for regeneration varies greatly among metazoans, yet little is known about the evolutionary processes leading to such different regeneration abilities. In particular, highly regenerative species such as planarians and cnidarians can regenerate the whole body from an amputated fragment; however, a common molecular basis, if any, among these species remains unclear. Here, we show that genes encoding Jumonji C (JmjC) domain-containing proteins are associated with high regeneration ability. We classified 132 fully sequenced metazoans into two groups with high or low regeneration abilities and identified 118 genes conserved in the high regenerative group that were lost in species in the low regeneration group during evolution. Ninety-six percent of them were JmjC domain-encoding genes. We denoted the candidate genes as high regenerative species-specific JmjC domain-encoding genes (HRJDs). We observed losses of HRJDs in Helobdella robusta, which lost its high regeneration ability during evolution based on phylogenetic analysis. By RNA sequencing analyses, we observed that HRJD orthologs were differentially expressed during regeneration in two Cnidarians, as well as Platyhelminthes and Urochordata, which are highly regenerative species. Furthermore, >50% of the head and tail parts of amputated planarians (Dugesia japonica) died during regeneration after RNA interference of HRJD orthologs. These results indicate that HRJD are strongly associated with a high regeneration ability in metazoans. HRJD paralogs regulate gene expression by histone demethylation; thus, HRJD may be related to epigenetic regulation controlling stem cell renewal and stem cell differentiation during regeneration. We propose that HRJD play a central role in epigenetic regulation during regeneration.


Beneficial impacts of fermented celery (Apium graveolens L.) juice on obesity prevention and gut microbiota modulation in high-fat diet fed mice.
Metabolic syndrome caused obesity has long been recognized as a risk of health. Celery and celery extracts have various medicinal properties, such as anti-diabetes and anti-inflammatory properties and blood glucose and serum lipid reduction. However, the effect of probiotic fermentation on celery juice and the association between fermented celery juice (FCJ) and obesity were unclear. This study aimed to evaluate the beneficial effects of FCJ on high-fat diet (HFD) induced obesity and related metabolic syndromes. C57BL/6 mice were randomly divided into six groups (n = 15 per group) fed either a normal diet (ND) or HFD with or without CJ/FCJ (10 g kg-1 day-1) by oral gavage for 12 weeks. Here we demonstrated that the probiotic fermentation of celery juice (CJ) could enhance the active ingredients in celery, such as total polyphenols, flavonoids, vitamin C and SOD. Compared to the slight improvement induced by CJ ingestion, FCJ intake significantly inhibited body weight gain, prevented dyslipidemia and hyperglycemia, and suppressed visceral fat accumulation. Furthermore, 16S rRNA sequencing analysis revealed that FCJ intake altered the composition of gut microbiota, increasing the ratio of Firmicutes/Bacteroidetes and the relative abundance of beneficial bacteria (Lactobacillus, Ruminococcaceae_UCG-014, Faecalibaculum and Blautia), and decreasing the relative abundance of harmful bacteria (Alloprevotella and Helicobacter). These findings suggest that FCJ can prevent HFD-induced obesity and become a novel gut microbiota modulator to prevent HFD-induced gut dysbiosis and obesity-related metabolic disorders.


T-Cell-Specific Loss of the PI-3-Kinase p110α Catalytic Subunit Results in Enhanced Cytokine Production and Antitumor Response.
Class IA phosphatidylinositol 3-kinase (PI3K) catalytic subunits p110α and p110δ are targets in cancer therapy expressed at high levels in T lymphocytes. The role of p110δ PI3K in normal or pathological immune responses is well established, yet the importance of p110α subunits in T cell-dependent immune responses is not clear. To address this problem, mice with p110α conditionally deleted in CD4+ and CD8+ T lymphocytes (p110α-/-ΔT) were used. p110α-/-ΔT mice show normal development of T cell subsets, but slightly reduced numbers of CD4+ T cells in the spleen. "In vitro," TCR/CD3 plus CD28 activation of naive CD4+ and CD8+ p110α-/-ΔT T cells showed enhanced effector function, particularly IFN-γ secretion, T-bet induction, and Akt, Erk, or P38 activation. Tfh derived from p110α-/-ΔT cells also have enhanced responses when compared to normal mice, and IL-2 expanded p110α-/-ΔT CD8+ T cells had enhanced levels of LAMP-1 and Granzyme B. By contrast, the expansion of p110α-/-ΔT iTreg cells was diminished. Also, p110α-/-ΔT mice had enhanced anti-keyhole limpet hemocyanin (KLH) IFN-γ, or IL-4 responses and IgG1 and IgG2b anti-KLH antibodies, using CFA or Alum as adjuvant, respectively. When compared to WT mice, p110α-/-ΔT mice inoculated with B16.F10 melanoma showed delayed tumor progression. The percentage of CD8+ T lymphocytes was higher and the percentage of Treg cells lower in the spleen of tumor-bearing p110α-/-ΔT mice. Also, IFN-γ production in tumor antigen-activated spleen cells was enhanced. Thus, PI3K p110α plays a significant role in antigen activation and differentiation of CD4+ and CD8+ T lymphocytes modulating antitumor immunity.


Tumor-related gene changes in immunosuppressive Syrian hamster cholangiocarcinoma.
The results of a previous study demonstrated that prednisolone enhanced cholangiocarcinogenesis. Therefore, to clarify molecular changes during immunosuppressive cholangiocarcinogenesis, Syrian hamsters were divided into 8 groups: uninfected controls; immunosuppressed Syrian hamsters using prednisolone (P); normal Syrian hamsters administered N-nitrosodimethylamine (ND); immunosuppressed Syrian hamsters administered N-nitrosodimethylamine (NDis); normal Syrian hamsters infected with Opisthorchis viverrini (OV); immunosuppressed Syrian hamsters infected with O. viverrini (OVis); normal Syrian hamsters infected with O. viverrini and administered N-nitrosodimethylamine (CCA); and immunosuppressed Syrian hamsters infected with O. viverrini and administered N-nitrosodimethylamine (CCAis). Syrian hamster livers were used for analysis of tumor-related gene expression and immunohistochemistry through cytokeratin 19 (CK19) and proliferating cell nuclear antigen (PCNA) staining. The tumor-related gene expression results show that CCAis groups at all time points exhibited upregulation of COX-2, IL-6, SOD1, CAT and iNOS and downregulation of p53, which correlated with the predominant expression of CK19 and PCNA in liver tissue. These results suggest that prednisolone enhances cholangiocarcinoma development, which was confirmed by molecular changes.


Further characterisation of the interaction of haloperidol metabolites with neurotransmitter transporters in rat neuronal cultures and in transfected COS-7 cells.
It has been proposed that the extrapyramidal symptoms such as tardive dyskinesia developed by patients on long-term haloperidol treatment may be the result of uptake of haloperidol metabolites into neurons via the monoamine neurotransmitter transporters followed by neurotoxic events, as occurs for MPP+, the pyridinium metabolite of MPTP. We recently showed that haloperidol and its metabolites are inhibitors of the human noradrenaline transporter (NAT), dopamine transporter (DAT) and serotonin transporter (SERT), and determined their Ki values for inhibition of the three transporters expressed in transfected COS-7 cells. In this study, we extended the investigation of these compounds to their inhibitory effects on DAT, SERT and the high affinity choline uptake (HACU) in neuronal cultures from embryonic rat brain, and investigated whether the compounds are substrates or non-transported inhibitors of the NAT, DAT and SERT in transfected COS-7 cells and DAT and SERT in the neuronal cultures. Haloperidol and its metabolites inhibited DAT, SERT and HACU in the neuronal cultures, indicating that they are not specific inhibitors of the monoamine neurotransmitter transporters. The ratio of the Ki values of the least and most potent inhibitors were found to be 2.8 for DAT, 24 for SERT and 7.6 for HACU. The compounds were more potent inhibitors of DAT and SERT in neuronal cultures than we found previously in transfected COS-7 cells. The question of whether the compounds are substrates or non-transported inhibitors of the monoamine transporters was investigated by determining whether they caused an increase in efflux of [3H]amine in transfected COS-7 cells or neuronal cultures preloaded with [3H]amine. Haloperidol metabolites were weak substrates for SERT, but not for NAT or DAT, in transporter-transfected COS-7 cells. In neuronal cultures, the metabolites appeared to be non-transported inhibitors or very weak substrates of DAT and SERT. Despite inhibition of the monoamine transporters by haloperidol and its metabolites, there is little evidence to support the proposal that these compounds are likely to cause neurotoxic effects via neuronal uptake using the monoamine transporters. The mechanisms of the side effects of haloperidol therapy, such as tardive dyskinesia, are still unclear, but are unlikely to depend on interactions of the drug or its metabolites with NAT, DAT or SERT.


Laing early-onset distal myopathy with subsarcolemmal hyaline bodies caused by a novel variant in the MYH7 gene.
Myopathies caused by MYH7 gene mutations are clinically and pathologically heterogeneous and, until recently, difficult to diagnose. The availability of NGS panels for hereditary neuromuscular diseases changed our insight regarding their frequency and allowed a better perception of the different phenotypes and morphological abnormalities associated. We present a male Portuguese patient with the classical phenotype of Laing early-onset distal myopathy (MPD1) beginning at 6 years of age, very slowly progressive, and with a mild to moderate impact on daily life by the age of 56. Muscle biopsy showed a myopathic pattern with hyaline bodies and cores. The NGS panel for structural myopathies identified a novel missense heterozygous variant, c.T4652C (p.Leu1551Pro), in the exon 34 of the MYH7 gene.


Sparse linear modeling of next-generation mRNA sequencing (RNA-Seq) data for isoform discovery and abundance estimation.
Since the inception of next-generation mRNA sequencing (RNA-Seq) technology, various attempts have been made to utilize RNA-Seq data in assembling full-length mRNA isoforms de novo and estimating abundance of isoforms. However, for genes with more than a few exons, the problem tends to be challenging and often involves identifiability issues in statistical modeling. We have developed a statistical method called "sparse linear modeling of RNA-Seq data for isoform discovery and abundance estimation" (SLIDE) that takes exon boundaries and RNA-Seq data as input to discern the set of mRNA isoforms that are most likely to present in an RNA-Seq sample. SLIDE is based on a linear model with a design matrix that models the sampling probability of RNA-Seq reads from different mRNA isoforms. To tackle the model unidentifiability issue, SLIDE uses a modified Lasso procedure for parameter estimation. Compared with deterministic isoform assembly algorithms (e.g., Cufflinks), SLIDE considers the stochastic aspects of RNA-Seq reads in exons from different isoforms and thus has increased power in detecting more novel isoforms. Another advantage of SLIDE is its flexibility of incorporating other transcriptomic data such as RACE, CAGE, and EST into its model to further increase isoform discovery accuracy. SLIDE can also work downstream of other RNA-Seq assembly algorithms to integrate newly discovered genes and exons. Besides isoform discovery, SLIDE sequentially uses the same linear model to estimate the abundance of discovered isoforms. Simulation and real data studies show that SLIDE performs as well as or better than major competitors in both isoform discovery and abundance estimation. The SLIDE software package is available at https://sites.google.com/site/jingyijli/SLIDE.zip.


Effect of chewing gums on plaque pH after a sucrose challenge.
The objective of this study was to determine whether sugarless chewing gums sweetened with different sweeteners differ in their ability to reduce an acidogenic response from a 10 percent sucrose-rinse challenge. Five commercially available chewing gums and two control regimens ("no gum" or paraffin) were tested using a plaque pH telemetry system. The gums were sweetened with sucrose, high-intensity sweeteners (aspartame, saccharin, or acesulfame-K), or a polyol (xylitol). Using a seven-period randomized block design, eight adult panelists were challenged with a 10 percent sucrose solution and then randomly used one of the test regimens during each of the seven test sessions. Each two-hour test session was divided into five periods: resting baseline (five minutes); sucrose rinse challenge (two minutes); postsucrose challenge (ten minutes); gum chewing (ten minutes); post gum chewing (ninety-three minutes). The factors analyzed were: the area of the curve (pH X Time) below pH 5.5, the minimum plaque pH attained, the changes in plaque pH over relevant intervals, and the length of time the plaque pH remained below pH 5.5. The various response variables showed a similar pattern of statistically significant differences. All of the sugarless gums were effective in significantly increasing plaque pH and in reducing the area under the curve after the sucrose challenge compared with "no gum" treatment. No statistically significant differences were noted among the sugarless gums. The response to sucrose gum was intermediate between sugarless gums and "no gum" but was not statistically different from "no gum" or three of the sugarless gums.(ABSTRACT TRUNCATED AT 250 WORDS)


[Viral hepatitis A: causes of lingering].
The examination covered 756 patients with viral hepatitis A (90 of them were followed up for 1.5 years) who had antigens and antibodies to hepatitis A, B, C, E and D viruses. It was found that: lingering viral hepatitis A may be caused by viral persistence marked by specific antibodies (IgM anti-HAV); lingering hepatitis followed by verification of chronic hepatitis is related to manifestation of previous hepatitis B and C; detection of IgM anti-HAV in patients with exacerbation of chronic hepatitis B, C and D (hepatitis D) evidences for the presence of mix-infection (acute hepatitis A in chronic hepatitis B, C or D).


Intimate partner violence: causes and prevention.
Unlike many health problems, there are few social and demographic characteristics that define risk groups for intimate partner violence. Poverty is the exception and increases risk through effects on conflict, women's power, and male identity. Violence is used as a strategy in conflict. Relationships full of conflict, and especially those in which conflicts occur about finances, jealousy, and women's gender role transgressions are more violent than peaceful relationships. Heavy alcohol consumption also increases risk of violence. Women who are more empowered educationally, economically, and socially are most protected, but below this high level the relation between empowerment and risk of violence is non-linear. Violence is frequently used to resolve a crisis of male identity, at times caused by poverty or an inability to control women. Risk of violence is greatest in societies where the use of violence in many situations is a socially-accepted norm. Primary preventive interventions should focus on improving the status of women and reducing norms of violence, poverty, and alcohol consumption.


Association of Serum Copeptin Levels with Patency of Infarct-Related Arteries in Patients with ST-Segment Elevation Myocardial Infarction.
Copeptin is widely used as a predictor of an adverse prognosis in many clinical conditions. Reduced antegrade coronary flow in an infarct-related artery (IRA) is associated with adverse clinical outcomes in patients with ST-segment elevation myocardial infarction (STEMI). The aim of this study was to investigate whether copeptin level on admission was associated with IRA patency in STEMI patients. A total of 88 patients were enrolled into the study and divided into two groups according to TIMI flow grade in the IRA before primary percutaneous coronary intervention. White blood cell count (p = 0.015), neutrophils (p = 0.047), N-terminal pro-brain natriuretic peptide (NTproBNP) (p < 0.001), copeptin (p < 0.001) and peak troponin I (p = 0.001) were significantly higher in the occluded IRA group with a significantly lower serum sodium level (p < 0.001). Age- and gender-adjusted multivariate analysis revealed that copeptin (OR = 1.970; p = 0.001), peak troponin I (1.055; p = 0.005) and NTproBNP (OR = 1.003; p = 0.010) were independent predictors of an occluded IRA. A copeptin cut-off value of > 6.8 ng/mL was found to predict an occluded IRA with a sensitivity of 80% and specificity of 100% (area under the curve: 0.917; p < 0.001). Performance ranking of the biomarkers that could predict an occluded IRA showed copeptin > peak troponin I = NTproBNP. Copeptin levels were higher in the patients with an occluded IRA and STEMI. Higher levels of copeptin predicted an occluded IRA in the patients with STEMI who were admitted to the emergency department during the first three hours of chest pain.


Signature of ray chaos in quasibound wave functions for a stadium-shaped dielectric cavity.
Light emission from a dielectric cavity with a stadium shape is studied in both ray and wave models. For a passive cavity mode with low loss, a remarkable correspondence is found between the phase space representation of a quasibound wave function and its counterpart distribution in the ray model. This result provides additional and more direct evidence for good ray-wave correspondence in low-loss modes previously observed at the level of far-field emission pattern comparisons.


Calcium channel blockers for preventing cardiomyopathy due to iron overload in people with transfusion-dependent beta thalassaemia.
Beta thalassaemia is a common inherited blood disorder. The need for frequent blood transfusions in this condition poses a difficult problem to healthcare systems. The most common cause of morbidity and mortality is cardiac dysfunction from iron overload. The use of iron chelation therapy has reduced the severity of systemic iron overload but specific, non-toxic treatment is required for removal of iron from the myocardium. To assess the effects of calcium channel blockers combined with standard iron chelation therapy in people with transfusion-dependent beta thalassaemia on the amount of iron deposited in the myocardium, on parameters of heart function, and on the incidence of severe heart failure or arrhythmias and related morbidity and mortality. We searched the Cochrane Haemoglobinopathies Trials Register, compiled from electronic database searches and handsearching of journals and conference abstract books. We also searched ongoing trials databases, and the reference lists of relevant articles and reviews.Date of last search: 24 February 2018. We included randomised controlled trials of calcium channel blockers combined with standard chelation therapy compared with standard chelation therapy alone or combined with placebo in people with transfusion-dependent beta thalassaemia. Two authors independently applied the inclusion criteria for the selection of trials. Two authors assessed the risk of bias of trials and extracted data and a third author verified these assessments. The authors used the GRADE system to assess the quality of the evidence. Two randomised controlled trials (n = 74) were included in the review; there were 35 participants in the amlodipine arms and 39 in the control arms. The mean age of participants was 24.4 years with a standard deviation of 8.5 years. There was comparable participation from both genders. Overall, the risk of bias in included trials was low. The quality of the evidence ranged across outcomes from low to high, but the evidence for most outcomes was judged to be low quality.Cardiac iron assessment, as measured by heart T2*, did not significantly improve in the amlodipine groups compared to the control groups at six or 12 months (low-quality evidence). However, myocardial iron concentration decreased significantly in the amlodipine groups compared to the control groups at both six months, mean difference -0.23 mg/g (95% confidence interval -0.07 to -0.39), and 12 months, mean difference -0.25 mg/g (95% confidence interval -0.44 to -0.05) (low-quality evidence). There were no significant differences between treatment and control groups in serum ferritin (low-quality evidence), liver T2* (low-quality evidence), liver iron content (low-quality evidence) and left ventricular ejection fraction (low-quality evidence). There were no serious adverse events reported in either trial; however, one trial (n = 59) reported mild adverse events, with no statistically significant difference between groups (low-quality evidence). The available evidence does not clearly suggest that the use of calcium channel blockers is associated with a reduction in myocardial iron in people with transfusion-dependent beta thalassaemia, although a potential for this was seen. There is a need for more long-term, multicentre trials to assess the efficacy and safety of calcium channel blockers for myocardial iron overload, especially in younger children. Future trials should be designed to compare commonly used iron chelation drugs with the addition of calcium channel blockers to investigate the potential interplay of these treatments. In addition, the role of baseline myocardial iron content in affecting the response to calcium channel blockers should be investigated. An analysis of the cost-effectiveness of the treatment is also required.


Potent Acetylcholinesterase Selective and Reversible Homodimeric Agent Based on Tacrine for Theranostics.
Acetylcholinesterase (AChE) has been an important biomarker for diagnosing Alzheimer's disease (AD), due to reduction in AChE activity in post-mortem brains of AD patients. A potent, selective, and reversible homodimeric inhibitor of AChE, 5-amino- N1, N3-bis(2-(1,2,3,4-tetrahydroacridin-9-ylamino)ethyl)isophthalamide (compound 4), was synthesized by using 9-alkyl(1,2,3,4-tetrahydroacridine) pharmacophore with appended functionality. In the present work, we report the synthesis of this bivalent inhibitor of AChE. The homodimeric ligand structure was designed and studied with molecular docking tools, which revealed its high affinity and interactions with active site gorge of AChE, which includes both catalytic active site (CAS) and peripheral active site (PAS). The IC50 value of this bivalent inhibitor for AChE and BuChE were 0.54 ± 0.06 and 32.49 ± 1.2 nM, respectively, with a selectivity ratio of 60.16 toward AChE. The designed ligand also showed potent inhibitory properties on PAS activity as well as on AChE-induced amyloid aggregation with low cytotoxicity on rat hippocampal neurons. The AFM images further corroborated the Aβ1-42 aggregation inhibition by compound 4 to an extent similar to bis(7)-tacrine. Moreover, the bivalent ligand was also proven to be of neurogenic potential due to its ability to induce S-phase post-treatment in rat hippocampal neuronal cells. On the basis of initial results, the agent could be further explored for its theranostic value clinically, which gives the possibility of tracing the AChE levels by molecular imaging techniques in correlation with progression of neurocognitive disorders like AD for better therapy response and patient management.


The cryopreservation and in vitro cultivation of larval Onchocerca volvulus.
A system developed in our laboratory for the in vitro cultivation of larval Onchocerca volvulus is being employed in a series of morphogenetic, physiologic, chemotherapeutic and immunologic investigations of this parasite. Because of the need for a large and readily available supply of living worms for this work, cryogenic techniques are being used for the long-term preservation of larval parasites collected in various endemic areas of Guatemala, C.A. To date, microfilariae have survived frozen storage in human cutaneous tissues (excised nodules and skin snips) for as long as 504 days, and viable larvae, in all stages of development have been recovered from the black fly vectors (Simulium ochraceum and S. metallicum) kept frozen for 396 days. That cryopreservation does not appear to affect these parasites adversely is indicated by the fact that microfilariae derived from frozen tissues do not differ from those obtained from fresh tissues on the basis of: 1) numbers and vigor of emergent microfilariae; 2) survival and morphogenesis of microfilariae during cultivation in vitro for 24 days; 3) glucose utilization during 72 hours of incubation; and 4) their incorporation of 3H-amino acids as determined after 18 hours of incubation. Details of methodology for cryopreservation and in vitro cultivation, together with resultant data, are presented herein.


Impact of climate change on potential distribution of Chinese caterpillar fungus (Ophiocordyceps sinensis) in Nepal Himalaya.
Climate change has already impacted ecosystems and species and substantial impacts of climate change in the future are expected. Species distribution modeling is widely used to map the current potential distribution of species as well as to model the impact of future climate change on distribution of species. Mapping current distribution is useful for conservation planning and understanding the change in distribution impacted by climate change is important for mitigation of future biodiversity losses. However, the current distribution of Chinese caterpillar fungus, a flagship species of the Himalaya with very high economic value, is unknown. Nor do we know the potential changes in suitable habitat of Chinese caterpillar fungus caused by future climate change. We used MaxEnt modeling to predict current distribution and changes in the future distributions of Chinese caterpillar fungus in three future climate change trajectories based on representative concentration pathways (RCPs: RCP 2.6, RCP 4.5, and RCP 6.0) in three different time periods (2030, 2050, and 2070) using species occurrence points, bioclimatic variables, and altitude. About 6.02% (8,989 km2) area of the Nepal Himalaya is suitable for Chinese caterpillar fungus habitat. Our model showed that across all future climate change trajectories over three different time periods, the area of predicted suitable habitat of Chinese caterpillar fungus would expand, with 0.11-4.87% expansion over current suitable habitat. Depending upon the representative concentration pathways, we observed both increase and decrease in average elevation of the suitable habitat range of the species.


Reliability, validity, incidence and impact of temporormandibular pain disorders in adolescents.
The first aim of this thesis was to investigate the prevalence of temporomandibular disorder (TMD) pain in adolescents an a Swedish county and whether there were differences in patient age and gender and in treatment given for TMD pain by dentists in Public Dental Service (PDS) clinics. The epidemiological variable TMD-S was introduced in the PDS in Ostergötland County, Sweden, in 2000 and is recorded for all adolescents aged 12-19 at the annual routine examination. Self-reported TMD pain in this investigation was based upon the response of the subjects to two questions: (1) Do you have pain in your temples, face, temporomandibular joint, or jaws once a week or more? and (2) Do you have pain when you open your mouth wide or chew, once a week or more? Dental records of 200 patients with TMD pain were randomly selected from the population to evaluate treatment given for TMD. Among 28,899 participating adolescents, 4.2% reported TMD pain. Prevalence increased with age, a significant difference was seen between boys and girls, and 34% of patients with TMD pain received TMD-relared treatment in dental clinics. The second aim was to evaluate the reliability and validity of self-reported TMD pain in 120 adolescents, 60 with self-reported TMD pain and 60 age- and sex-matched controls without TMD pain. All adolescents were examined twice at a PDS clinic. At the first examination, self-reported TMD pain was recorded for each patient. At the second examination, a clinical examination was made blind to the patients' self-report of pain symptoms, after which self-reported TMD pain was again recorded. The clinical examination was based upon the Research Diagnostic Criteria for TMD (RDC/TMD). Test-retest reliability of 0.83 (kappa) was found for the two questions. Sensitivity was 0.98 and specificity 0.90 for assessments made on the same day and 0.96 and 0.83, respectively, for assessments made 2-4 weeks apart. The third aim was to evaluate incidence, by age and gender, and temporal patterns of TMD pain in adolescents. This 3-year longitudinal study was carried out at all PDS clinics from 2000 to 2003. All individuals aged 12-19 years in the county who visited the clinics for annual examinations were eligible for the study. Overall, the annual incidence of TMD pain among 2255 participating adolescents was 2.9%. Incidence among girls (4.5% was significantly higher than in boys (1.3%). Incidence increased with age in girls and boys, although less so in boys. These adolescents were re-examined annually for 3 years, and a fluctuating pattern of TMD pain was common. The fourth and final aim was to investigate gender and age differences in pain behavior, jaw function, and psychosocial status in adolescents with self-reported TMD pain. A postal questionnaire was sent to 350 consecutive patients with self-reported TMD pain and 350 healthy age- and sex-matched individuals aged 12-19 years 2-4 weeks after their annual dental examination. The groups were divided into younger (age 12-15) and older (age 16-19) groups. The TMD and control groups differed significantly in most variables related to pain characteristics and psychosocial and behavioral factors. Multiple pain sites were significantly more common in the TMD than in the control group, but there were no gender differences. For adolescents reporting pain once a week or more, no gender differences were seen in pain intensities. Jaw function limitation, depression scores, and perceived need for TMD treatment were significantly higher overall in girls than in boys. Almost one-third of older girls, compared to one out of ten older boys, reported school absences and analgesic consumption because of their TMD pain. Older girls had significantly higher Graded Chronic Pain Scale scores than older boys. In conclusion, TMD pain increases with increasing age in adolescents and is more common in girls than in boys. A fluctuating pain pattern can be seen. TMD-S, with two self-report questions, has very good reliability and validity, and can be recommended for screening adolescents for TMD pain. TMD pain seems to have a greater Impact on girls than boys. particularly in ages 16-19 years.


Primary radiotherapy with endobronchial high-dose-rate brachytherapy boost for inoperable lung cancer: long-term results.
To retrospectively evaluate the outcome of patients with inoperable non-small-cell lung cancer treated with primary external beam radiotherapy combined with high-dose-rate endobronchial brachytherapy boost. Between 1988 and 2005, 35 patients with non-small-cell lung cancer (stage I-III) ineligible for surgical resection and/or chemotherapy, were primarily treated with external beam radiotherapy with a median total dose of 50 Gy (range, 46-60). A median of 3 fractions high-dose-rate endobronchial brachytherapy was applied as a boost after external beam radiotherapy, the median total dose was 15 Gy (range, 8-20). High-dose-rate endobronchial brachytherapy was carried out with iridium-192 sources (370 GBq) and prescribed to 1 cm distance from the source axis. With a median follow-up of 26 months from the first fraction of high-dose-rate endobronchial brachytherapy, the 1-, 2- and 5-year overall (local progression-free) survival rates were 76% (76%), 61% (57%) and 28% (42%), respectively. Complete or partial remission rates 6 to 8 weeks after treatment were 57% and 17%, respectively. Significant prognostic favorable factors were a complete remission 6-8 weeks after treatment and a negative nodal status. In patients without mediastinal node involvement, a long-term local control could be achieved with 56% 5-year local progression-free survival. Common Toxicity Criteria grade 3 toxicities were hemoptysis (n = 2) and necrosis (n = 1). One fatal hemoptysis occurred in combination with a local tumor recurrence. The combination of external beam radiotherapy with high-dose-rate endobronchial brachytherapy boost is an effective primary treatment with acceptable toxicity in patients with non-small-cell lung cancer ineligible for surgical resection and/or chemotherapy.


InChI in the wild: an assessment of InChIKey searching in Google.
While chemical databases can be queried using the InChI string and InChIKey (IK) the latter was designed for open-web searching. It is becoming increasingly effective for this since more sources enhance crawling of their websites by the Googlebot and consequent IK indexing. Searchers who use Google as an adjunct to database access may be less familiar with the advantages of using the IK as explored in this review. As an example, the IK for atorvastatin retrieves ~200 low-redundancy links from a Google search in 0.3 of a second. These include most major databases and a very low false-positive rate. Results encompass less familiar but potentially useful sources and can be extended to isomer capture by using just the skeleton layer of the IK. Google Advanced Search can be used to filter large result sets. Image searching with the IK is also effective and complementary to open-web queries. Results can be particularly useful for less-common structures as exemplified by a major metabolite of atorvastatin giving only three hits. Testing also demonstrated document-to-document and document-to-database joins via structure matching. The necessary generation of an IK from chemical names can be accomplished using open tools and resources for patents, papers, abstracts or other text sources. Active global sharing of local IK-linked information can be accomplished via surfacing in open laboratory notebooks, blogs, Twitter, figshare and other routes. While information-rich chemistry (e.g. approved drugs) can exhibit swamping and redundancy effects, the much smaller IK result sets for link-poor structures become a transformative first-pass option. The IK indexing has therefore turned Google into a de-facto open global chemical information hub by merging links to most significant sources, including over 50 million PubChem and ChemSpider records. The simplicity, specificity and speed of matching make it a useful option for biologists or others less familiar with chemical searching. However, compared to rigorously maintained major databases, users need to be circumspect about the consistency of Google results and provenance of retrieved links. In addition, community engagement may be necessary to ameliorate possible future degradation of utility.


[Neurology in the court of the Learned King].
During the reign of Alfonso X of Castile one of the most flourishing cultural periods of European history was reached. The so-called Learned King contributed to this development by promoting the use of Castilian as the language of scientific discovery instead of Latin, and favoring the recovery of Arabic origin manuscripts in the Toledo Translators' School. Where the wise men from the "three cultures" (Christian, Islamic and Jewish) worked together in harmony to perpetuate classical knowledge. The aim of this paper is to analyze neurological pathology in the period concerned. The Learned King suffered from several neurologic syndromes including delirium and coma. He also showed chronic maxillary sinusitis with orbital involvement, trigeminal neuralgia and visual loss. Furthermore, in the "Cantigas" he described miraculous treatments of several disorders including seizures, leprous neuropathy, dementia, rabies, and ergotism.


Cutaneous Manifestations of Non-Celiac Gluten Sensitivity: Clinical Histological and Immunopathological Features.
The dermatological manifestations associated with intestinal diseases are becoming more frequent, especially now when new clinical entities, such as Non-Celiac Gluten Sensitivity (NCGS), are identified. The existence of this new entity is still debated. However, many patients with diagnosed NCGS that present intestinal manifestations have skin lesions that need appropriate characterization. We involved 17 patients affected by NCGS with non-specific cutaneous manifestations who got much better after a gluten free diet. For a histopathological and immunopathological evaluation, two skin samples from each patient and their clinical data were collected. The median age of the 17 enrolled patients affected by NCGS was 36 years and 76% of them were females. On the extensor surfaces of upper and lower limbs in particular, they all presented very itchy dermatological manifestations morphologically similar to eczema, psoriasis or dermatitis herpetiformis. This similarity was also confirmed histologically, but the immunopathological analysis showed the prevalence of deposits of C3 along the dermo-epidermal junction with a microgranular/granular pattern (82%). The exact characterization of new clinical entities such as Cutaneous Gluten Sensitivity and NCGS is an important objective both for diagnostic and therapeutic purposes, since these are patients who actually benefit from a GFD (Gluten Free Diet) and who do not adopt it only for fashion.


Melanin-Concentrating Hormone acts through hypothalamic kappa opioid system and p70S6K to stimulate acute food intake.
Melanin-Concentrating Hormone (MCH) is one of the most relevant orexigenic factors specifically located in the lateral hypothalamic area (LHA), with its physiological relevance demonstrated in studies using several genetically manipulated mice models. However, the central mechanisms controlling MCH-induced hyperphagia remain largely uncharacterized. Here, we show that central injection of MCH in mice deficient for kappa opoid receptor (k-OR) failed to stimulate feeding. To determine the hypothalamic area responsible for this MCH/k-OR interaction, we performed virogenetic studies and found that downregulation of k-OR by adeno-associated viruses (shOprk1-AAV) in LHA, but not in other hypothalamic nuclei, was sufficient to block MCH-induced food intake. Next, we sought to investigate the molecular signaling pathway within the LHA that mediates acute central MCH stimulation of food intake. We found that MCH activates k-OR and that increased levels of phosphorylated extracellular signal regulated kinase (ERK) are associated with downregulation of phospho-S6 Ribosomal Protein. This effect was prevented when a pharmacological inhibitor of k-OR was co-administered with MCH. Finally, the specific activation of the direct upstream regulator of S6 (p70S6K) in the LHA attenuated MCH-stimulated food consumption. Our results reveal that lateral hypothalamic k-OR system modulates the orexigenic action of MCH via the p70S6K/S6 pathway.