Fonds de Recherche du Québec - Santé

BACKGROUND: A population-based phylogenetic approach was used to characterize human immunodeficiency virus (HIV)-transmission dynamics in Quebec. METHODS: HIV-1 pol sequences included primary HIV infections (PHIs; <6 months after seroconversion) from the Quebec PHI cohort (1998-2005; n=215) and the provincial genotyping program (2001-2005; n=481). Phylogenetic analysis determined sequence interrelationships among unique PHIs (n=593) and infections from untreated (n=135) and treated (n=660) chronically infected (CI) potential transmitter populations (2001-2005). Clinical features, risk factors, and drug resistance for clustered and nonclustered transmission events were ascertained. RESULTS: Viruses from 49.4% (293/593) of PHIs cosegregated into 75 transmission chains with 2-17 transmissions/cluster. Half of the clusters included 2.7+/-0.8 (mean+/-SD) transmissions, whereas the remainder had 8.8+/-3.5 transmissions. Maximum periods for onward transmission in clusters were 15.2+/-9.5 months. Coclustering of untreated and treated CIs with PHIs were infrequent (6.2% and 4.8%, respectively). The ages, viremia, and risk factors were similar for clustered and nonclustered transmission events. Low prevalence of drug resistance in PHI supported amplified transmissions at early stages. CONCLUSIONS: Early infection accounts for approximately half of onward transmissions in this urban North American study. Therapy at early stages of disease may prevent onward HIV transmission.

RATIONALE: Dexmedetomidine is associated with less delirium than benzodiazepines and better sleep architecture than either benzodiazepines or propofol; its effect on delirium and sleep when administered at night to patients requiring sedation remains unclear. OBJECTIVES: To determine if nocturnal dexmedetomidine prevents delirium and improves sleep in critically ill adults. METHODS: This two-center, double-blind, placebo-controlled trial randomized 100 delirium-free critically ill adults receiving sedatives to receive nocturnal (9:30 p.m. to 6:15 a.m.) intravenous dexmedetomidine (0.2 μg/kg/h, titrated by 0.1 μg /kg/h every 15 min until a goal Richmond Agitation and Sedation Scale score of -1 or maximum rate of 0.7 μg/kg/h was reached) or placebo until ICU discharge. During study infusions, all sedatives were halved; opioids were unchanged. Delirium was assessed using the Intensive Care Delirium Screening Checklist every 12 hours throughout the ICU admission. Sleep was evaluated each morning by the Leeds Sleep Evaluation Questionnaire. MEASUREMENTS AND MAIN RESULTS: Nocturnal dexmedetomidine (vs. placebo) was associated with a greater proportion of patients who remained delirium-free during the ICU stay (dexmedetomidine [40 (80%) of 50 patients] vs. placebo [27 (54%) of 50 patients]; relative risk, 0.44; 95% confidence interval, 0.23-0.82; P = 0.006). The average Leeds Sleep Evaluation Questionnaire score was similar (mean difference, 0.02; 95% confidence interval, 0.42-1.92) between the 34 dexmedetomidine (average seven assessments per patient) and 30 placebo (six per patient) group patients able to provide one or more assessments. Incidence of hypotension, bradycardia, or both did not differ significantly between groups. CONCLUSIONS: Nocturnal administration of low-dose dexmedetomidine in critically ill adults reduces the incidence of delirium during the ICU stay; patient-reported sleep quality appears unchanged. Clinical trial registered with www.clinicaltrials.gov (NCT01791296).

Motor learning changes the activity of cortical motor and subcortical areas of the brain, but does learning affect sensory systems as well? We examined in humans the effects of motor learning using fMRI measures of functional connectivity under resting conditions and found persistent changes in networks involving both motor and somatosensory areas of the brain. We developed a technique that allows us to distinguish changes in functional connectivity that can be attributed to motor learning from those that are related to perceptual changes that occur in conjunction with learning. Using this technique, we identified a new network in motor learning involving second somatosensory cortex, ventral premotor cortex, and supplementary motor cortex whose activation is specifically related to perceptual changes that occur in conjunction with motor learning. We also found changes in a network comprising cerebellar cortex, primary motor cortex, and dorsal premotor cortex that were linked to the motor aspects of learning. In each network, we observed highly reliable linear relationships between neuroplastic changes and behavioral measures of either motor learning or perceptual function. Motor learning thus results in functionally specific changes to distinct resting-state networks in the brain.

Glaucoma, a major cause of blindness worldwide, is a neurodegenerative optic neuropathy in which vision loss is caused by loss of retinal ganglion cells (RGCs). To better define the pathways mediating RGC death and identify targets for the development of neuroprotective drugs, we developed a high-throughput RNA interference screen with primary RGCs and used it to screen the full mouse kinome. The screen identified dual leucine zipper kinase (DLK) as a key neuroprotective target in RGCs. In cultured RGCs, DLK signaling is both necessary and sufficient for cell death. DLK undergoes robust posttranscriptional up-regulation in response to axonal injury in vitro and in vivo. Using a conditional knockout approach, we confirmed that DLK is required for RGC JNK activation and cell death in a rodent model of optic neuropathy. In addition, tozasertib, a small molecule protein kinase inhibitor with activity against DLK, protects RGCs from cell death in rodent glaucoma and traumatic optic neuropathy models. Together, our results establish a previously undescribed drug/drug target combination in glaucoma, identify an early marker of RGC injury, and provide a starting point for the development of more specific neuroprotective DLK inhibitors for the treatment of glaucoma, nonglaucomatous forms of optic neuropathy, and perhaps other CNS neurodegenerations.

The recovery of locomotion, following interactive training with graded weight support, in the adult spinal cat has led to the proposal that removal of body weight may be a therapeutic tool in human gait retraining. There would be benefits, however, in knowing normal responses of humans to partial weight bearing before applying this strategy to patients. In this study, 10 nondisabled male subjects walked on a treadmill while 0%, 30%, 50%, and 70% of their body weight was supported by a modified climbing harness. To dissociate the changes attributable to walking speed from those attributable to body weight, each subject walked at the specified body-weight-support (BWS) levels and at full weight bearing (FWB) at the same speed. Simultaneously, electromyographic data from the right leg muscles, footswitch signals, and video recording of joint motion were collected. The FWB and BWS gaits appeared similar, except at the highest level of BWS studied (ie, 70% of BWS). Significant differences among other BWS and FWB trials at comparable speeds included decreases in percentage of stance, percentage of total double-limb support time, and maximum hip and knee flexor swing angle. Other adaptations to BWS were a reduction in the mean burst amplitude of the muscles that are active during stance and an increase in the mean burst amplitude of the tibialis anterior muscle. The possible implications of this new gait retraining strategy for patients with neurological impairment are discussed. [Finch L, Barbeau H, Arsenault B. Influence of body weight support on normal human gait: development of a gait retraining strategy.

OBJECTIVE: The purpose of this study was to determine whether the New Injury Severity Score (NISS) is a better predictor of mortality than the Injury Severity Score (ISS) in general and in subgroups according to age, penetrating trauma, and body region injured. METHODS: The study population consisted of 24,263 patients from three urban Level I trauma centers in the province of Quebec, Canada. Discrimination and calibration of NISS and ISS models were compared using receiver operator characteristic (ROC) curves and Hosmer-Lemeshow statistics. RESULTS: NISS showed better discrimination than ISS (area under the ROC curve = 0.827 vs. 0.819; p = 0.0006) and improved calibration (Hosmer-Leme-show = 62 vs. 112). The advantage of the NISS over the ISS was particularly evident among patients with head/neck injuries (area under the ROC curve = 0.819 vs. 0.784; p < 0.0001; Hosmer-Lemeshow = 59 vs. 350). CONCLUSION: The NISS is a more accurate predictor of in-hospital death than the ISS and should be chosen over the ISS for case-mix control in trauma research, especially in certain subpopulations such as head/neck-injured patients.

BACKGROUND: While minimalist running shoes may have an influence on running biomechanics and on the incidence of overuse injuries, the term "minimalist" is currently used without standardisation. The objectives of this study were to reach a consensus on a standard definition of minimalist running shoes, and to develop and validate a rating scale that could be used to determine the degree of minimalism of running shoes, the Minimalist Index (MI). METHODS: For this modified Delphi study, 42 experts from 11 countries completed four electronic questionnaires on an optimal definition of minimalist shoes and on elements to include within the MI. Once MI was developed following consensus, 85 participants subjectively ranked randomly assigned footwear models from the most to the least minimalist and rated their degree of minimalism using visual analog scales (VAS), before evaluating the same footwear models using MI. A subsample of thirty participants reassessed the same shoes on another occasion. Construct validity and inter- and intra-rater reliability (intraclass correlation coefficients [ICC]; Gwet's AC1) of MI were evaluated. RESULTS: The following definition of minimalist shoes was agreed upon by 95 % of participants: "Footwear providing minimal interference with the natural movement of the foot due to its high flexibility, low heel to toe drop, weight and stack height, and the absence of motion control and stability devices". Characteristics to be included in MI were weight, flexibility, heel to toe drop, stack height and motion control/stability devices, each subscale carrying equal weighing (20 %) on final score. Total MI score was highly correlated with VAS (r = 0.91). A significant rank effect (p < 0.001) confirmed the MI's discriminative validity. Excellent intra- and inter-rater reliability was found for total MI score (ICC = 0.84-0.99) and for weight, stack height, heel to toe drop and flexibility subscales (AC1 = 0.82-0.99), while good inter-rater reliability was found for technologies (AC1 = 0.73). CONCLUSION: This standardised definition of minimalist shoes developed by an international panel of experts will improve future research on minimalist shoes and clinical recommendations. MI's adequate validity and reliability will allow distinguishing running shoes based on their degree of minimalism, and may help to decrease injuries related to footwear transition.

The provocative concentrations of inhaled methacholine that cause 6% (PC6) and 20% (PC20) falls in forced expiratory volume in one second (FEV1) were assessed in a population of 100 nonsmoking persons, equally distributed for sex, who ranged uniformly from 20 to 60 yr of age. These subjects had no respiratory symptoms, rhinitis, atopic history, or familial history of asthma. Single twofold dilutions of methacholine from 2 to 128 mg/ml were used; 81 and 34 subjects, respectively, showed PC6 and PC20 values less than 128 mg/ml. Eight subjects had PC20 values less than 16 mg/ml. In these subjects, the test had a good reproducibility (r = 0.92) when we repeated it, and serial measurements of peak expiratory flow rates did not suggest asthma. The fact that PC6 was related, although loosely, to baseline FEV, FEV/FVC, and forced expiratory flow during the middle half of the FVC (FEF) and that 4 of the 8 subjects with PC20 values less than 16 mg/ml had lower values of FEF might suggest that responsiveness to methacholine is partially linked with baseline airway caliber.

OBJECTIVE: Osteoarthritis (OA) is accompanied by subchondral bone sclerosis. The present study was undertaken to determine whether osteoblast-like cells in patients with OA show an abnormal phenotype that could contribute to this sclerosis. METHODS: Explants and primary in vitro osteoblast-like cell cultures were prepared from subchondral bone specimens from OA patients or from bone removed at autopsy from individuals showing no signs of OA or metabolic bone disease. We measured the abundance and activity of urokinase plasminogen activator (uPA), and the levels of PA inhibitor (PAI-1) and insulin-like growth factor 1 (IGF-1) in conditioned media from both explants and osteoblast-like cells. The expression of osteoblast phenotypic biomarkers was also evaluated. RESULTS: OA explants showed increased levels and activity of uPA, no changes in PAI-1 abundance, and increases in IGF-1 release, as compared with preparations from normal individuals. In vitro primary osteoblast-like cells showed results similar to the ex vivo findings for uPA, PAI-1, and IGF-1. Primary OA osteoblast-like cells also expressed higher alkaline phosphatase activity and osteocalcin release than normal cells, both under basal conditions and with 1,25(OH)2D3 (1,25-dihydroxyvitamin D) stimulation. Conversely, OA osteoblast-like cells showed blunted cAMP synthesis in response to human parathyroid hormone and prostaglandin E2 in contrast to the finding with normal osteoblast-like cells, a result that could not be attributed to altered adenylate cyclase activity. CONCLUSION: Ex vivo and in vitro results indicate similar altered activities of OA osteoblasts as compared with normal cells. This suggests that an altered phenotype of subchondral osteoblasts may be a contributing factor in human OA.

OBJECTIVE: First, to validate an ultrasonographic measure of the acromio-humeral distance (AHD); second, to compare the AHD variation during active abduction in patients with shoulder impingement syndrome (SIS) and healthy subjects; and third, to evaluate the relationship between functional status and AHD variations before and after rehabilitation in SIS subjects. DESIGN: This study has 3 components: (1) a reliability study, (2) a case-control study, and (3) a preliminary pretreatment/posttreatment clinical trial. SETTING: Primary care hospital setting. PARTICIPANTS: Seven SIS patients and 13 healthy subjects. INTERVENTIONS: For the clinical trial, the SIS subjects participated in 12 sessions of a rehabilitation program over 4 weeks. MAIN OUTCOME MEASURES: First, intraclass correlation coefficient for interobserver reliability; second, AHD measured at 0 degrees, 45 degrees, and 60 degrees of active abduction; and third, Western Ontario Rotator Cuff Index. RESULTS: Intraclass correlation coefficient for interobserver reliability ranged from 0.86 to 0.92 for the 3 shoulder positions. A significant reduction of the AHD was found within groups between rest and active abduction (P < 0.05). Comparison of AHD between groups was not statistically different (P = 0.06; beta < 0.80). In pre-post rehabilitation analysis, improvement of the Western Ontario Rotator Cuff Index score was positively correlated to the reduction of the AHD narrowing as the arm was abducted (r = 0.86; P = 0.01). CONCLUSIONS: The ultrasound measure of AHD is reliable and sensitive. Although a distinct pattern of AHD variation in SIS patients could not be confirmed, a strong positive relationship was found between the reduction of AHD narrowing and functional improvement following rehabilitation. Ultrasound measurement of AHD might help identify SIS patients who will benefit from rehabilitation.

The routine use of antibiotics in agriculture has contributed to an increase in drug-resistant bacterial pathogens in animals that can potentially be transmitted to humans. In 2000, the World Health Organization identified resistance to antibiotics as one of the most significant global threats to public health and recommended that the use of antibiotics as additives in animal feed be phased out or terminated, particularly those used to treat human infections. Research is currently being carried out to identify alternative antimicrobial compounds for use in animal production. A number of studies, mostly in vitro, have provided evidence indicating that bacteriocins, which are antimicrobial peptides of bacterial origin, may be promising alternatives to conventional antibiotics in poultry and swine production. This review provides an update on bacteriocins and their potential for use in the poultry and swine industries.

Enamelin is critical for proper dental enamel formation, and defects in the human enamelin gene cause autosomal dominant amelogenesis imperfecta. We used gene targeting to generate a knock-in mouse carrying a null allele of enamelin (Enam) that has a lacZ reporter gene replacing the Enam translation initiation site and gene sequences through exon 7. Correct targeting of the transgene was confirmed by Southern blotting and PCR analyses. No enamelin protein could be detected by Western blotting in the Enam-null mice. Histochemical 5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside (X-gal) staining demonstrated ameloblast-specific expression of enamelin. The enamel of the Enam+/- mice was nearly normal in the maxillary incisors, but the mandibular incisors were discolored and tended to wear rapidly where they contacted the maxillary incisors. The Enam-/- mice showed no true enamel. Radiography, microcomputed tomography, and light and scanning electron microscopy were used to document changes in the enamel of Enam-/- mice but did not discern any perturbations of bone, dentin, or any other tissue besides the enamel layer. Although a thick layer of enamel proteins covered normal-appearing dentin of unerupted teeth, von Kossa staining revealed almost a complete absence of mineral formation in this protein layer. However, a thin, highly irregular, mineralized crust covered the dentin on erupted teeth, apparently arising from the formation and fusion of small mineralization foci (calcospherites) in the deeper part of the accumulated enamel protein layer. These results demonstrate ameloblast-specific expression of enamelin and reveal that enamelin is essential for proper enamel matrix organization and mineralization. Enamelin is critical for proper dental enamel formation, and defects in the human enamelin gene cause autosomal dominant amelogenesis imperfecta. We used gene targeting to generate a knock-in mouse carrying a null allele of enamelin (Enam) that has a lacZ reporter gene replacing the Enam translation initiation site and gene sequences through exon 7. Correct targeting of the transgene was confirmed by Southern blotting and PCR analyses. No enamelin protein could be detected by Western blotting in the Enam-null mice. Histochemical 5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside (X-gal) staining demonstrated ameloblast-specific expression of enamelin. The enamel of the Enam+/- mice was nearly normal in the maxillary incisors, but the mandibular incisors were discolored and tended to wear rapidly where they contacted the maxillary incisors. The Enam-/- mice showed no true enamel. Radiography, microcomputed tomography, and light and scanning electron microscopy were used to document changes in the enamel of Enam-/- mice but did not discern any perturbations of bone, dentin, or any other tissue besides the enamel layer. Although a thick layer of enamel proteins covered normal-appearing dentin of unerupted teeth, von Kossa staining revealed almost a complete absence of mineral formation in this protein layer. However, a thin, highly irregular, mineralized crust covered the dentin on erupted teeth, apparently arising from the formation and fusion of small mineralization foci (calcospherites) in the deeper part of the accumulated enamel protein layer. These results demonstrate ameloblast-specific expression of enamelin and reveal that enamelin is essential for proper enamel matrix organization and mineralization. Enamel crystals have a unique shape and organization. They are many times longer than they are wide and are organized into rods (prisms), each comprising about 10,000 individual crystallites (1Daculsi G. Kerebel B. J. Ultrastruct. Res. 1978; 65: 163-172Crossref PubMed Scopus (133) Google Scholar, 2Nanci, A. (ed) (2003) in Ten Cate's Oral Histology Development, Structure, and Function, 6th Ed., pp. 218-224, Mosby, St. Louis, MOGoogle Scholar). Each rod is the product of a single ameloblast in which crystals are formed under the control of specialized enamel proteins secreted by ameloblasts (3Fincham A.G. Moradian-Oldak J. Simmer J.P. J. Struct. Biol. 1999; 126: 270-299Crossref PubMed Scopus (496) Google Scholar). These proteins assemble immediately subjacent to the secretory pole of the ameloblast cell membrane such that mineralization occurs immediately at this cell-matrix interface (4Ronnholm E. J. Ultrastruct. Res. 1962; 6: 249-303Crossref PubMed Scopus (137) Google Scholar). Because enamel crystals grow longer at this mineralization front, the ameloblasts are displaced away from the growing tooth as the enamel layer as a whole thickens. Once the enamel crystals are fully elongated and the enamel layer has reached its final thickness, the secretion of enamel proteins is terminated or greatly reduced, accumulated extracellular proteins are degraded and reabsorbed, and the crystals grow in width and thickness until adjacent crystals come into contact. Thus, the secretion of enamel proteins is associated with the early part of crystal formation, when the crystals are growing primarily in length and the growing crystal tips are in close proximity to the secretory surface of the ameloblast. There are three major secretory stage enamel proteins: amelogenin (5Snead M.L. Zeichner-David M. Chandra T. Robson K.J. Woo S.L. Slavkin H.C. Proc. Natl. Acad. Sci. U. S. A. 1983; 80: 7254-7258Crossref PubMed Scopus (93) Google Scholar, 6Snead M.L. Lau E.C. Zeichner-David M. Fincham A.G. Woo S.L. Slavkin H.C. Biochem. Biophys. Res. Commun. 1985; 129: 812-818Crossref PubMed Scopus (177) Google Scholar), ameloblastin (7Krebsbach P.H. Lee S.K. Matsuki Y. Kozak C.A. Yamada K. Yamada Y. J. Biol. Chem. 1996; 271: 4431-4435Abstract Full Text Full Text PDF PubMed Scopus (261) Google Scholar), and enamelin (8Hu C.-C. Fukae M. Uchida T. Qian Q. Zhang C.H. Ryu T. Y. M. Simmer J.P. J. Res. PubMed Scopus Google Scholar, Qian Q. Zhang C.H. Simmer J.P. J. Res. PubMed Scopus Google Scholar). to the secretory gene K. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar), and are critical for proper dental enamel is the secreted enamel protein and is from the or the and the E.C. Slavkin H.C. M.L. PubMed Scopus Google Scholar, E.C. P.H. K. J. Google Scholar). cause amelogenesis in that the enamel layer of the J. A. PubMed Scopus Google Scholar, T. Simmer J.P. PubMed Scopus Google Scholar). mice amelogenesis B. G. E. T. T. S. G. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). Although amelogenin expression has detected in many the Y. J. Sci. PubMed Scopus Google Scholar, A. A. Y. B. Y. E. E. B. S. J.P. J. Sci. PubMed Scopus Google Scholar), no defects in have in with amelogenesis imperfecta. No of amelogenesis by human ameloblastin have but mice enamel S. T. B. T. G. P.H. A. Yamada Y. J. Biol. PubMed Scopus Google Scholar). enamelin gene cause autosomal dominant amelogenesis K. PubMed Scopus Google Scholar, B. K. Simmer J.P. J. Res. PubMed Scopus Google Scholar, Y. Oral Biol. PubMed Scopus Google Scholar). a that a single allele a of amelogenesis defects in the enamel layer Ryu S. G. E. J. PubMed Scopus Google Scholar, E. G. Ryu J. Res. PubMed Scopus Google Scholar). in the mouse enamelin gene have with the and Enam have the site in exon and K. Y. J. A. A. K. J. T. Y. T. S. T. PubMed Scopus Google Scholar, M. U. M. J. Res. PubMed Scopus Google Scholar). The enamel a and enamel surface in mice and enamel in the null to the three secretory are secreted into the enamel matrix used matrix microcomputed scanning electron and Simmer J.P. J. H.C. E.C. 1996; PubMed Scopus Google Scholar, J.P. Fukae M. T. Y. Uchida T. J. H.C. M. J. Res. PubMed Scopus Google Scholar). is secreted in the early stage with and is in the stage Zhang S. Simmer J.P. J. Oral Sci. PubMed Scopus Google Scholar, J. J. and J. C.-C. in of the on and pp. Scholar). in the cause autosomal amelogenesis in Ryu S. J. PubMed Scopus Google Scholar, Simmer J.P. J. PubMed Scopus Google Scholar, Ryu M. E. J. Res. PubMed Scopus Google Scholar). mice have a enamel that is than normal and to the surface J. Y. Simmer J.P. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, E. Lee J. Res. PubMed Scopus Google Scholar). expression to be to Ryu J. Simmer J.P. H.C. Res. Scopus Google Scholar), is in many but is in and in K. Proc. Natl. Acad. Sci. U. S. A. 1999; PubMed Scopus Google Scholar, Zhang Y. Ryu Simmer J.P. PubMed Scopus Google Scholar, S. M. and E. Chem. Scholar). is that and are for proper dental enamel formation, but they are not in and for about of of amelogenesis Simmer J.P. J. Sci. PubMed Scopus Google Scholar). the of amelogenesis of the proteins in the enamel layer of have and is that not of the major extracellular have J. and J. (2003) in of the on and pp. Scholar). The secretion of with the enamel proteins that enamelin and ameloblastin be from These enamel proteins have and other M. T. Uchida T. M. Res. 1996; PubMed Scopus Google Scholar, C.-C. Fukae M. Uchida T. Qian Q. Zhang C.H. Ryu T. Y. M. Simmer J.P. J. Res. PubMed Scopus Google Scholar), and to generate with the have to in any T. Y. P.H. Simmer J.P. J. Res. PubMed Scopus Google Scholar). is to the secretory stage extracellular matrix and the formation of enamel crystals in into the dental enamel formation and to the normal and of Enam have used gene targeting to normal Enam replacing the Enam with that of a reporter in mice. We have demonstrated that Enam is and by ameloblasts and that Enam-null mice not enamel of a complete to at the in the of a mineralization at the secretory surface of the ameloblast. were and by the on and of at the of of the targeting with Western a a lacZ reporter with a mouse of a a into the unique site in the of the The was by that could be by The Enam was by PCR as and into the site of The Enam in and in was into the The final targeting was by and by with and into by of in the that was for by Southern were and into The were to and the were for by Southern three The was with and and the and the knock-in were The was with and and the and the knock-in were was with and and to for targeting the for the was of the were with mice to the and mice were that The or absence of a or was by PCR or and or were by PCR of by for Enam and was a control a Enam at demonstrated the of at Enam lacZ at demonstrated the of at knock-in The mice were for which a mice were on for and in a and three mouse enamelin were from for of and were from mice for each of the three and a of was at the of the of on the were from mice to of in and for staining M. A. Biol. 1999; PubMed Scopus Google Scholar, Y. Y. J. A. PubMed Google Scholar, Y. Zhang Y. 129: PubMed Google Scholar). The were in for to to the of the mice and of the tissue with three times for and in for to to the of the mice PubMed Scopus Google Scholar). The were three times in and in were at thickness, for in three times in and at for in staining and Biochem. PubMed Scopus Google Scholar). were and in for with and under a or a light were a and and of mouse were and the were and on in a The incisors and were tissue under a and the and enamel were and in on and for each enamelin were and with of and at and the were by and with The were a and and of protein was in of Each was by on three was with the were to and with proteins for Western analyses. membrane was amelogenin J.P. Lau E.C. T. M. Zeichner-David M. M.L. Slavkin H.C. Fincham A.G. PubMed Scopus Google the other membrane was the enamelin and with the were with to was detected by and enamelin was detected by of mandibular incisors from each of the three at were Oral Biol. PubMed Scopus Google Scholar, J. Res. PubMed Scopus Google Scholar). at the at for for the of was immediately the for dental with a was used to a in in the mandibular and the enamel and dentin of the was by a and The mice were and the were and at The incisors were on a and the the on the and the on the was with a three times The the in the and the on the was by the of that the were the of was of and Enam+/- mice were from and a at from three mice from each of the three were and under was on at the of the from three of each The was at and at were a by a to the a of scanning was for each which of were on surface to a of the to of the and of the were a of of at microscopy of at the of the unerupted was on and mice at of were with in and in were with a on to Kossa staining for mineral was by to the and to light for were for tissue and cell were a on of at from mice for each of the three were with a at the of the and with were with a to and with a scanning electron in the electron at of of and at mandibular from and knock-in mice were in in and in a at and were and and with and a of Enamel at and mice at of were and for tissue and were of and in in The were for at The and enamel the incisors were and the enamel were with The enamel layer on each maxillary and mandibular was with a into a of from the the Each was in a small and at The were to and on a to the of the enamel Each was to a and and at for The were and each enamel was to the of the and any mineral in each enamel H.C. J. Res. PubMed Scopus Google Scholar). of this the protein of a was as the the final The mineral by in each was by the from were from a of maxillary and mandibular incisors or incisors and in of three of enamel early and nearly of for that normal of were by Enamelin and in the of enamelin on crystal crystals were in a G. Biochem. J. PubMed Scopus Google in the of or enamelin. were from the and to with a and a highly J. Res. PubMed Scopus Google Scholar). of the used gene targeting to generate a mouse carrying a null allele of enamelin (Enam) that has a lacZ reporter gene into the of the Enam the targeting in Enam are to as Enam-/- or as to the of Enam expression or the of expression the of the enamelin The lacZ which was to a mouse and into the Enam exon replacing the Enam translation initiation of the Enam through was that the lacZ enamelin the part of Enam through The sequences for the were with and by mice with mice. 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The is that stage ameloblasts are for staining but for in is the of the protein than Enam to in the generate expression of the protein has Enam expression was detected by and by no other cell the highly The close the lacZ staining in the mice and normal ameloblast-specific expression enamelin as by in Zhang Simmer J.P. J. Oral Sci. PubMed Scopus Google that the knock-in was Western the that Enam was by with the targeting is by the results of Western analyses. Enamel proteins were from incisors and and of proteins from and Enam-/- mice were by and Western Enamelin protein was detected in from and Enam+/- mice but not from the enamelin null mice when the proteins from the Enam-/- were in times the that could be detected in Enam+/- no enamelin protein could be detected and of the mouse at changes in tooth shape and the and Enam mice in the Enam-/- mice enamel and the Enam+/- mice defects that are The mandibular incisors of the Enam+/- mice are in the erupted and the maxillary incisors are from normal to The mandibular enamel the part of the is apparently rapidly a wear on the Enamel is on the of the Enam+/- mice The of wear is on the Enam-/- which are and at the The maxillary incisors of Enam+/- mice normal to have in the enamel with and the wear on the tips of were not or as as on the maxillary incisors of The mandibular incisors of Enam+/- in with of and and the of enamel the of the incisors. wear on were with enamel and dentin the in to the on erupted of the in tips that a shape than the shape of incisors. of enamel were on the maxillary and the mandibular incisors of Enam-/- mice. These a and showed dentin the and of the The erupted of the incisors were with a layer of that and in The of the were with a layer of and showed of were in the and of showed the mandibular with and in to the of the and There to be tissue in the under the There was for tissue in to the of the incisors. was in the enamel the but was in the the the enamel were than was and from the and of were in the stage at the and the There was a small on the incisors where the and Enam+/- and Enam-/- mice could not be from in of of of and were on and the and mice did not any in or when with There was no mice of the three in a of the of and to be in critical mineralization and a in a The that the targeting Enam in lacZ was by Southern PCR and the dental in the Enam+/- and Enam-/- mice. The results that was no in and the the expression of Enam by the that the targeting a of the dental incisors as they and the for the enamel layer to J. C.A. Res. PubMed Scopus Google Scholar). from with the tooth its of S. M. Res. PubMed Scopus Google Scholar). Although enamelin is not to a in tooth from wear or a of to in changes in the of mandibular were in mice the enamel and adjacent and the the a of was not to the enamel in this of the of the enamel layer in the Enam a was through the and and the were on to in the and enamelin mice were The for mandibular incisors of mice was The for the Enam-null mice was The for the mice was three were than for mouse mandibular incisors Oral Biol. PubMed Scopus Google Scholar), of the of and The mandibular for the Enam+/- mice to in the the maxillary and mandibular incisors are maxillary incisors of Enam+/- mice normal The incisors of Enam+/- in a of and with enamel and dentin at the the mandibular incisors of the Enam+/- mice maxillary incisors the wear of the mandibular incisors and this for The in the and Enam-/- mice are not to cause in the of mineralization of the mice under demonstrated that the of the mouse mandibular dental at from the to the Enam+/- to the Enam-/- mice with the in tooth mineralization in the The mineralized of the Enam-/- mice were and than in the that the Enam-null mice enamel and at the of the mandibular and in the the and that the mineralized enamel layer in the Enam-/- mice is to be detected or enamel is in the and Enam+/- mice. of mandibular through the and of by and that could be used to the mineralized tissue and tooth of the Enam mice. The revealed a of the mineralized tissue a of mineralized in the and from Enam-/- mice to the and Enam+/- no were in and The and a or of dental enamel in the Enam-/- mice. No in or tooth could be the and Enam+/- mice. mandibular of the mice were von Kossa staining and which mineralized to but not or was that the secretory stage enamel layer of the Enam-/- mice was von for small that in the No von Kossa staining was at the mineralization front, which is the secretory of the membrane of the ameloblasts the surface of the enamel The secretory stage enamel of the and Enam+/- mice the enamel of the mice did not as as that of the mice. that enamel proteins in the enamel but the of enamel of the mineral dentin in the Enam-/- mice is true which by the of crystal 10,000 each at the mineralization that grow in width and thickness to the enamel (1Daculsi G. Kerebel B. J. Ultrastruct. Res. 1978; 65: 163-172Crossref PubMed Scopus (133) Google Scholar). electron of mouse and incisors revealed early enamel layer with the of and enamel S. J. 1996; Google Scholar). the accumulated enamel matrix of the Enam-null mice showed of enamel rods that the and enamel of at showed enamel in the stage of enamel rods were in the and Enam+/- mice and but were from the Enam-null mice. the Enam-/- mice the accumulated enamel proteins were and a layer of mineral the The surface of the was and to or The in Enam+/- mice but that the enamel on than and were of and of the enamel surface that were the of the and of the and of of enamel of length were from the to the of mouse incisors. Each was and its Each was which the and the of the The protein is the the The mineral the The were by major and for the and Enam-/- mice The and mineral of enamel on maxillary incisors from the and Enam+/- mice were the mandibular incisors of Enam+/- mice and mineral that were almost of for enamel at in mice and The mineral to protein in enamel on mandibular incisors of the Enam+/- mice was normal the secretory normal early when enamel crystals growing in and normal at a when the enamel in mice was almost to with a were for the mineral by was to enamel from the erupted of teeth, of normal with enamel on mandibular incisors of mice not We were to of the length of the maxillary and mandibular incisors in Enam-null mice. The maxillary and mandibular incisors showed for and mineral the of the incisors and about of the for mice. The mineral to protein were of normal and the mineral by at stage of was the nearly enamel in mice was mineral by The mineral to protein and mineral by for the Enam-null mice were almost to for normal not Enamelin and of the enamelin protein is degraded and from the enamel with the of a product that to about of enamel protein T. T. E.C. Fukae M. M. PubMed Scopus Google Scholar). crystals were in in the of enamelin. showed that crystal length was in the absence of enamelin and with protein to at crystal did not with enamelin and at and enamelin the length of crystals in not the crystal that enamelin is the protein the enamel matrix of The proteins and are in and are to and Because of its and from the the is the used for enamel proteins Y. P.H. Simmer J.P. PubMed Scopus Google Scholar). enamelin is secreted as a that is rapidly by in the extracellular Y. Oral Biol. PubMed Scopus Google Scholar). The secreted protein is from its to generate and but not and are the enamel surface M. T. Uchida T. M. Res. 1996; PubMed Scopus Google Scholar, T. Y. Fukae M. Y. K. M. Simmer J.P. Uchida T. Res. PubMed Scopus Google Scholar). and of into ameloblasts the of the matrix that mineralization. The of enamelin is that for the enamelin at the mineralization where they are and no is deeper in the enamel (8Hu C.-C. Fukae M. Uchida T. Qian Q. Zhang C.H. Ryu T. Y. M. Simmer J.P. J. Res. PubMed Scopus Google Scholar). to other of the enamelin protein the thickness of enamel T. T. E.C. Fukae M. M. PubMed Scopus Google Scholar, T. T. Fukae M. M. PubMed Scopus (93) Google Scholar, T. T. Fukae M. M. Yamada M. K. S. PubMed Scopus Google Scholar). The of the enamelin protein the at the mineralization has that enamelin is critical for enamel crystal the of a thick layer of enamel proteins in the extracellular secretory ameloblasts in the Enam-/- no mineralization occurs at the mineralization that enamelin is a critical of the mineralization that or the of enamel crystals J.P. Fincham A.G. Oral Biol. 6: PubMed Scopus Google Scholar). the enamel crystals that at the are to at the mineralization (4Ronnholm E. J. Ultrastruct. Res. 1962; 6: 249-303Crossref PubMed Scopus (137) Google Scholar, A. (ed) in Ten Cate's Oral Histology Development, Structure, and Function, Ed., pp. Mosby, St. Louis, MOGoogle and the of enamel crystal formation to be in the Enam-/- be that the initiation and of enamel crystallites occurs by the or a of the and mineralization of of the of a that is in The small of that dentin in the Enam-/- mice to be than true is of not and is the product of ameloblasts the mineral to protein and mineral by are to for We that is no true enamel the dentin in the Enam-/- mice. The mineral that by a than for true enamel. to the enamel protein layer as that and a crust the dentin that These the of the mineralization at the secretory surface of the ameloblast membrane as to the formation of true enamel. The shape of the enamel crystals and organization into rods is at the mineralization and is the enamelin as the enamel layer and in of the enamelin have for enamel crystals and in the the enamel with the of ameloblastin T. Y. Fukae M. Y. K. M. Simmer J.P. Uchida T. Res. PubMed Scopus Google Scholar). The enamelin product is the enamelin that the crystallites enamel the thickness of secretory stage enamel T. T. E.C. Fukae M. M. PubMed Scopus Google Scholar, T. T. Fukae M. M. PubMed Scopus (93) Google Scholar). and three Y. PubMed Scopus Google Scholar, Y. T. Fukae M. M. Res. PubMed Scopus Google Scholar). The enamelin not in with amelogenin and J. in and and pp. but for to Y. T. S. Simmer J.P. Fukae M. Oral Biol. PubMed Scopus Google Scholar). part of enamelin to its to by Y. Fukae M. T. Simmer J.P. J. Oral Sci. PubMed Scopus Google Scholar), which is the in secretory stage enamel Fincham A.G. Zhang Qian Q. Simmer J.P. J. Res. 1999; PubMed Scopus Google Scholar). the enamelin the length but not the of crystals in These results that the enamelin a in enamel crystallites in The reporter in of enamelin the highly of Enam enamelin is by any cell other than be at to secretory highly of expression with which is by ameloblasts but be detected in many other to that enamelin expression in other be and amelogenin expression be mouse was that a Enam of the translation initiation site expression of the mouse enamelin gene at than normal a Enam was by in the and not by The and for that the gene expression of enamelin J. J. and J. C.-C. J. in Scholar). have and Enam knock-in The results that enamelin is by ameloblasts and that enamelin expression is for proper of the mineralization associated with the ameloblast secretory enamelin is no true enamel. Enamelin defects are in as for enamelin is to the enamel formation tooth We of of and and of for and was by at and of the knock-in and of the mouse were by with

OBJECTIVE: To record the prevalence, extent, cost, and satisfaction with use of alternative medicine practices by patients with fibromyalgia syndrome (FMS), compared to control rheumatology patients. METHODS: An interviewer-based questionnaire was administered to 221 consecutive rheumatology patients and 80 FMS patients. RESULTS: Alternative medicine interventions were currently being used extensively by rheumatology patients overall, and by FMS patients in particular. All categories of alternative practices were used more often by FMS patients, compared to controls, including overall use 91% versus 63% (P = 0.0001), over-the-counter products 70% versus 54% (NS), spiritual practices 48% versus 37% (NS), and alternative practitioners 26% versus 12% (P = 0.003), respectively. Two-thirds of patients using alternative medicine practices were concurrently using multiple interventions. Patient satisfaction ratings were highest for spiritual interventions. CONCLUSIONS: Alternative medicine practices were currently being used by almost all FMS patients. This observation might indicate that traditional medical therapies are inadequate in providing symptomatic relief to FMS patients.

OBJECTIVE: To determine the prevalence and clinical correlations of an anomaly consisting of electroencephalographic (EEG) waves within the alpha frequency band during non-rapid eye movement (NREM) sleep in patients with fibromyalgia, and to evaluate the alpha NREM sleep anomaly as a predictor of response to amitriptyline. METHODS: Twenty-two patients with fibromyalgia were studied in a 2-month, double-blind, crossover trial of amitriptyline (25 mg/day) versus placebo. Nocturnal EEGs were conducted on 2 consecutive nights at baseline and at the end of each 2-month treatment period. RESULTS: Six patients (27%) had a clinical response to amitriptyline, while none responded to placebo (P = 0.02). Treatment with amitriptyline or placebo did not result in any changes in the alpha ratings during NREM sleep. Only 8 patients (36%) exhibited the alpha NREM sleep anomaly at baseline. Those patients reported more sleep difficulty, but otherwise were clinically indistinguishable from those without this EEG sleep anomaly. Lower baseline alpha NREM sleep ratings were seen in responders to amitriptyline than in nonresponders, but these differences did not reach statistical significance. CONCLUSION: The alpha NREM sleep anomaly is present in only a small proportion of patients with fibromyalgia. It does not correlate with disease severity nor is it affected by treatment with amitriptyline. A larger sample size will be needed to adequately assess the value of this sleep anomaly in predicting the response to amitriptyline.

We sequenced genes coding for components of the SNARE complex (STX1A, VAMP2, SNAP25) and their regulatory proteins (STXBP1/Munc18-1, SYT1), which are essential for neurotransmission, in 95 patients with idiopathic mental retardation. We identified de novo mutations in STXBP1 (nonsense, p.R388X; splicing, c.169+1G>A) in two patients with severe mental retardation and nonsyndromic epilepsy. Reverse transcriptase polymerase chain reaction and sequencing showed that the splicing mutation creates a stop codon downstream of exon-3. No de novo or deleterious mutations in STXBP1 were found in 190 control subjects, or in 142 autistic patients. These results suggest that STXBP1 disruption is associated with autosomal dominant mental retardation and nonsyndromic epilepsy.

Kingdon's multiple‐streams framework, which emerged in the mid‐1980s, today forms one of the indispensable analytical frameworks for understanding public policy agenda‐setting. However, it is only in the context of wealthy countries that this approach has been validated for setting the agenda of national and international policies. This article reports the results of empirical research in an African state studying the transferability of a threefold theoretical innovation. The question under consideration is whether the multiple‐streams framework is useful for examining public policy implementation at the local level and in the context of a low income country. The research findings confirm the premise that the multiple‐streams framework can be extended and can lead to the formulation of several theoretical propositions.

Innovation has the potential to improve the quality of care and health service delivery, but maximising the reach and impact of innovation to achieve large-scale health system transformation remains understudied. Interest is growing in three processes of the innovation journey within health systems, namely the spread, sustainability and scale-up (3S) of innovation. Recent reviews examine what we know about these processes. However, there is little research on how to support and operationalise the 3S. This study aims to improve our understanding of the 3S of healthcare innovations. We focus specifically on the definitions of the 3S, the mechanisms that underpin them, and the conditions that either enable or limit their potential. We conducted a scoping review, systematically investigating six bibliographic databases to search, screen and select relevant literature on the 3S of healthcare innovations. We screened 641 papers, then completed a full-text review of 112 identified as relevant based on title and abstract. A total of 24 papers were retained for analysis. Data were extracted and synthesised through descriptive and inductive thematic analysis. From this, we develop a framework of actionable guidance for health system actors aiming to leverage the 3S of innovation across five key areas of focus, as follows: (1) focus on the why, (2) focus on perceived-value and feasibility, (3) focus on what people do, rather than what they should be doing, (4) focus on creating a dialogue between policy and delivery, and (5) focus on inclusivity and capacity building. While there is no standardised approach to foster the 3S of healthcare innovations, a variety of practical frameworks and tools exist to support stakeholders along this journey.

BALB/c mice immunized with the phospholipid, cardiolipin, produced anti-cardiolipin and anti-DNA antibodies. Seven hybridomas derived from spleen cells of the cardiolipin-immunized mice produced cardiolipin-binding monoclonal antibodies that also bound to the polynucleotides DNA, poly(dT), and poly(I). The seven cardiolipin-induced monoclonal antibodies shared idiotypic determinants with a high frequency idiotypic marker of spontaneously expressed anti-DNA autoantibodies of lupus-prone MRL-lpr/lpr mice. The monoclonal antibodies presumably bound to phosphodiester phosphate groups that occur in both polynucleotides and phospholipids. The results imply that production of anti-DNA autoantibodies does not require immunization by DNA.

Our aim was to examine the potential incremental impact of vaccinating boys against human papillomavirus (HPV) on vaccine-type infection in females and males, using an individual-based HPV transmission-dynamic model. Under base assumptions (vaccine efficacy = 99%, duration of protection = 20 years, coverage = 70%), vaccinating 12-year-old boys, in addition to girls, resulted in an incremental reduction in HPV-16/18 (HPV-6/11) incidence over 70 years of 16% (3%) in females and 23% (4%) in males. The benefit of vaccinating boys decreased with improved vaccination coverage in girls. Given the important predicted herd immunity impact of vaccinating girls under moderate to high vaccine coverage, the potential incremental gains of vaccinating boys are limited.

BACKGROUND: We estimated the lifetime medical costs attributable to sexually transmitted infections (STIs) acquired in 2018, including sexually acquired human immunodeficiency virus (HIV). METHODS: We estimated the lifetime medical costs of infections acquired in 2018 in the United States for 8 STIs: chlamydia, gonorrhea, trichomoniasis, syphilis, genital herpes, human papillomavirus (HPV), hepatitis B, and HIV. We limited our analysis to lifetime medical costs incurred for treatment of STIs and for treatment of related sequelae; we did not include other costs, such as STI prevention. For each STI, except HPV, we calculated the lifetime medical cost by multiplying the estimated number of incident infections in 2018 by the estimated lifetime cost per infection. For HPV, we calculated the lifetime cost based on the projected lifetime incidence of health outcomes attributed to HPV infections acquired in 2018. Future costs were discounted at 3% annually. RESULTS: Incident STIs in 2018 imposed an estimated $15.9 billion (25th-75th percentile: $14.9-16.9 billion) in discounted, lifetime direct medical costs (2019 US dollars). Most of this cost was due to sexually acquired HIV ($13.7 billion) and HPV ($0.8 billion). STIs in women accounted for about one fourth of the cost of incident STIs when including HIV, but about three fourths when excluding HIV. STIs among 15- to 24-year-olds accounted for $4.2 billion (26%) of the cost of incident STIs. CONCLUSIONS: Incident STIs continue to impose a considerable lifetime medical cost burden in the United States. These results can inform health economic analyses to promote the use of cost-effective STI prevention interventions to reduce this burden.