Rate and loudness manipulations in dysarthria: acoustic and perceptual findings.
Both rate reduction and increased loudness reportedly are associated with an increase in the size of the articulatory-acoustic working space and improved acoustic distinctiveness for speakers with dysarthria. Improved intelligibility also has been reported. Few studies have directly compared rate and loudness effects for speakers with dysarthria, however, although rate reduction and increasing vocal loudness are common treatment techniques. In the current study, 15 speakers with dysarthria secondary to multiple sclerosis, 12 speakers with dysarthria secondary to Parkinson's disease (PD), and 15 healthy controls read a passage in habitual, loud, and slow conditions. Rate and loudness variations were elicited using magnitude production. Acoustic measures included articulatory rate, sound pressure level, vowel space area, first moment difference measures, and F2 trajectory characteristics for diphthongs. Ten listeners scaled intelligibility for reading passages produced by the speakers with dysarthria. Relationships between intelligibility estimates and acoustic measures were determined by regression analysis. All speaker groups reduced articulatory rate for the slow condition and increased vocal intensity for the loud condition, relative to the habitual condition. Vowel acoustic distinctiveness, as indexed by vowel space area, was maximized in the slow condition, but stop consonant acoustic distinctiveness, as indexed by first moment difference measures, was maximized in the loud condition. F2 slope measures for diphthongs were not consistently affected by rate or loudness. Scaled intelligibility for speakers with PD also improved in the loud condition relative to both the habitual and slow conditions. Intelligibility estimates for speakers with dysarthria, however, were not strongly related to acoustic measures of supraglottal behavior. Findings are compared with previous studies, and hypotheses for future treatment studies are discussed.


Follicular T-cell lymphoma mimicking lymphocyte-rich classic Hodgkin lymphoma: a case report of a diagnostic pitfall.
Follicular T-cell lymphoma (FTCL), one of the nodal T-cell lymphomas with T follicular helper (TFH) phenotype, is an uncommon disease. The diagnosis of FTCL is challenging on the distinction from the morphological mimics mostly exemplified by follicular lymphoma. Here, we described a case of FTCL that mimicked lymphocyte-rich classic Hodgkin lymphoma (LRCHL). A 47-year-old male presented with cervical lymphadenopathy. The biopsy specimen demonstrated nodular lymphoid proliferation, which included scattered CD30+ CD15- CD20- PAX5 weakly+ Hodgkin and Reed-Sternberg (HRS)-like cells and a rich distribution of CD3+ CD4+ PD1+ T-cells. Epstein Barr virus was not detected in HRS-like cells, but it was detected in a small proportion of the scattered lymphocytes. The large cells were also negative for programmed cell death ligand 1, which appeared to be coincidental as described in our previous report of LRCHL. However, flow cytometry showed a CD3- CD4+ T-cell population that constituted 37.4% of all gated lymphocytes. A PCR analysis showed a clonal T-cell receptor-gamma gene rearrangement, but not a clonal immunoglobulin heavy chain gene rearrangement, and showed RHOA G17V mutation. The constellation of these findings led us to revise the diagnosis to FTCL. This result indicated that our case belonged to a relatively indolent subgroup of nodal peripheral T-cell lymphoma of TFH phenotype, which affects patients ≤60 years old, recently proposed by our group. This case report expands our understanding of the morphologic spectrum of FTCL and its clinicopathologic significance.


Macrocyclic Encapsulation in a Non-fused Tetrathiophene Acceptor for Efficient Organic Solar Cells with High Short-Circuit Current Density.
Non-fullerene acceptors have shown great promise for organic solar cells (OSCs). However, challenges in achieving high efficiency molecular system with conformational unicity and effective molecular stacking remain. In this study, we present a new design of non-fused tetrathiophene acceptor R4T-1 via employing the encapsulation of tetrathiophene with macrocyclic ring. The single crystal structure analysis reveals that cyclic alkyl side chains can perfectly encapsulate the central part of molecule and generate a conformational stable and planar molecular backbone. Whereas, the control 4T-5 without the encapsulation restriction displays cis- and twisted conformation. As a result, R4T-1 based OSCs achieved an outstanding power conversion efficiency (PCE) exceeding 15.10 % with a high short-circuit current density (Jsc ) of 25.48 mA/cm2 , which is significantly improved by ≈30 % in relative to that of the control. Our findings demonstrate that the macrocyclic encapsulation strategy could assist fully non-fused electron acceptors (FNEAs) to achieve a high photovoltaic performance and pave a new way for FNEAs design.


Quantitative nuclear proteomics identifies mTOR regulation of DNA damage response.
Cellular nutritional and energy status regulates a wide range of nuclear processes important for cell growth, survival, and metabolic homeostasis. Mammalian target of rapamycin (mTOR) plays a key role in the cellular responses to nutrients. However, the nuclear processes governed by mTOR have not been clearly defined. Using isobaric peptide tagging coupled with linear ion trap mass spectrometry, we performed quantitative proteomics analysis to identify nuclear processes in human cells under control of mTOR. Within 3 h of inhibiting mTOR with rapamycin in HeLa cells, we observed down-regulation of nuclear abundance of many proteins involved in translation and RNA modification. Unexpectedly, mTOR inhibition also down-regulated several proteins functioning in chromosomal integrity and up-regulated those involved in DNA damage responses (DDRs) such as 53BP1. Consistent with these proteomic changes and DDR activation, mTOR inhibition enhanced interaction between 53BP1 and p53 and increased phosphorylation of ataxia telangiectasia mutated (ATM) kinase substrates. ATM substrate phosphorylation was also induced by inhibiting protein synthesis and suppressed by inhibiting proteasomal activity, suggesting that mTOR inhibition reduces steady-state (abundance) levels of proteins that function in cellular pathways of DDR activation. Finally, rapamycin-induced changes led to increased survival after radiation exposure in HeLa cells. These findings reveal a novel functional link between mTOR and DDR pathways in the nucleus potentially operating as a survival mechanism against unfavorable growth conditions.


Neurotoxicity induced by antiepileptic drugs in cultured hippocampal neurons: a comparative study between carbamazepine, oxcarbazepine, and two new putative antiepileptic drugs, BIA 2-024 and BIA 2-093.
Newly designed antiepileptic drugs (AEDs) are being evaluated for their efficacy in preventing seizures and for their toxic profiles. We investigated and compared the toxic effects of two dibenz[b,f]azepine derivatives with anticonvulsant activity, 10,11-dihydro-10-hydroxyimino-5H-dibenz[b,f]azepine-5-carboxamide (BIA2-024) and (S)-(-)-10-acetoxy-10,11-dihydro-5H-dibenz[b,f] azepine-5-carboxamide (BIA2-093), with the structurally related compounds carbamazepine (CBZ) and oxcarbazepine (OXC), both in current use for the treatment of epilepsy. Primary rat hippocampal neurons were used to evaluate neuronal morphology and biochemical changes induced by the AEDs used in this study. Immunocytochemical staining against MAP-2 was used to evaluate neuronal morphology. Reactive oxygen species (ROS) and changes in mitochondrial membrane potential (Psim) were measured by fluorescence techniques. Intracellular adenosine triphosphate (ATP) levels were quantified by high-performance liquid chromatography (HPLC). Hippocampal neurons treated for 24 h with CBZ or OXC (300 microM) showed degeneration and swelling of neurites, but this effect was not observed in neurons treated with BIA 2-024 or BIA 2-093 (300 microM). ROS production also was increased in neurons treated with OXC, but not in neurons treated with the other AEDs. ATP levels were significantly decreased only in neurons treated with OXC, although the energy charge was not altered. Furthermore, OXC led to a decrease of Psim. In all parameters assayed, OXC was more toxic than the other AEDs used. Because the new putative AEDs have previously been shown to have an efficacy in preventing seizures similar to that of CBZ and OXC, and are less toxic to neuronal cells, they may be considered as alternatives to the current available therapies for the treatment of epilepsy.


Natural and induced ovarian synchrony in golden lion tamarins (Leontopithecus rosalia).
Ovarian cycle synchrony was assessed in spontaneously cycling female golden lion tamarins by monitoring longitudinal (16 mo) urinary steroid metabolite (estrone conjugates; pregnanediol-3 alpha-glucuronide, PdG) excretion in four pairs (n = 8) of females isolated from males. The overall mean ovarian cycle duration was 18.5 +/- 0.3 days (n = 136 cycles; mean range, 15.7-21.0 days), and there was no evidence of reproductive seasonality. Laparoscopic ovarian examinations confirmed that cyclic fluctuations in urinary steroid metabolite excretion were temporally associated with the formation and demise of corpora lutea. Evaluation of ovarian synchronization tested the null hypothesis that urinary hormone cycles were expressed randomly relative to those of cagemates or other females housed in separate cages but within close proximity. Natural ovarian synchrony (expressed as the mean difference in ovarian cycle onset) between cagemates (4.1 +/- 0.4 days) and among noncagemates (4.2 +/- 0.2 days) did not differ (p > 0.05) from a random ovarian cycle distribution. Two trials also were conducted to evaluate the efficacy of the prostaglandin (PG) F2 alpha analogue, cloprostenol, for artificially synchronizing ovarian cycles. Induced ovarian synchrony was not achieved with a single 0.8-microgram i.m. injection of cloprostenol. However, doubling the cloprostenol dose (1.6 micrograms) caused a rapid decrease in mean urinary PdG (p < 0.05) within 2 days, and synchronous ovulation was demonstrated by an increase (p < 0.01) in mean urinary PdG 10 days after cloprostenol administration. In summary, females housed in pairs, in the absence of males, exhibit spontaneous, year-round ovarian cycles with no evidence of among-female ovarian synchrony. Results also suggest that this New World primate has a reduced sensitivity to cloprostenol (compared to common marmosets) but that a single, midcycle cloprostenol injection of 1.6 micrograms effectively induces luteolysis and synchronous ovulation.


LRRK2 G2019S-induced mitochondrial DNA damage is LRRK2 kinase dependent and inhibition restores mtDNA integrity in Parkinson's disease.
Mutations in leucine-rich repeat kinase 2 (LRRK2) are associated with increased risk for developing Parkinson's disease (PD). Previously, we found that LRRK2 G2019S mutation carriers have increased mitochondrial DNA (mtDNA) damage and after zinc finger nuclease-mediated gene mutation correction, mtDNA damage was no longer detectable. While the mtDNA damage phenotype can be unambiguously attributed to the LRRK2 G2019S mutation, the underlying mechanism(s) is unknown. Here, we examine the role of LRRK2 kinase function in LRRK2 G2019S-mediated mtDNA damage, using both genetic and pharmacological approaches in cultured neurons and PD patient-derived cells. Expression of LRRK2 G2019S induced mtDNA damage in primary rat midbrain neurons, but not in cortical neuronal cultures. In contrast, the expression of LRRK2 wild type or LRRK2 D1994A mutant (kinase dead) had no effect on mtDNA damage in either midbrain or cortical neuronal cultures. In addition, human LRRK2 G2019S patient-derived lymphoblastoid cell lines (LCL) demonstrated increased mtDNA damage relative to age-matched controls. Importantly, treatment of LRRK2 G2019S expressing midbrain neurons or patient-derived LRRK2 G2019S LCLs with the LRRK2 kinase inhibitor GNE-7915, either prevented or restored mtDNA damage to control levels. These findings support the hypothesis that LRRK2 G2019S-induced mtDNA damage is LRRK2 kinase activity dependent, uncovering a novel pathological role for this kinase. Blocking or reversing mtDNA damage via LRRK2 kinase inhibition or other therapeutic approaches may be useful to slow PD-associated pathology.


Postvasectomy semen analysis: are men following up?
Many family physicians perform outpatient vasectomies in their office. Postvasectomy semen analysis (PVSA) is critical to establish the success of this sterilization procedure. We investigated the compliance rate of our patients with the PVSA over a 10-year period. To determine compliance rates of men who have undergone vasectomy for recommended PVSA, a retrospective chart review in a private family practice clinic was performed. The records of all patients who underwent vasectomy from 1991 to 2001 were reviewed. Patients are instructed to return at 6 weeks, 3 months, and 1 year for semen specimen evaluation to determine vasectomy success. Records of 551 patients were reviewed. The age of the patient at the time of the vasectomy and number of PVSAs were evaluated. PVSA compliance rates were also correlated with age. In total, 233 (42%) men did not return for 6-week, 3-month, or 1-year PVSA. Of 551 men, 318 (58%) returned for 6-week PVSA, and 138 (25%) returned for 3-month PVSA. Only 44 (8%) of the 551 eligible for a 1-year analysis returned for PVSA. Compliance with instructions to men undergoing vasectomy to return for PVSA is low both from the perspective of this study, as well as other studies evaluated. Older men are more likely to return for PVSA.


An exaggerated epinephrine-adrenergic receptor signaling impairs uterine decidualization in mice.
Our understanding of the relationship between stress-derived epinephrine and early pregnancy failure remains incomplete. Here, we explored the effect of epinephrine exposure on early pregnancy and pseudopregnancy in mice. Increased expression of adrenergic receptors Adra1b, Adra2b and Adrb2 was observed during decidualization and post-implantation embryogenesis was delayed or survival impaired. Epinephrine treatment also impaired decidualization in both the gravid and pseudopregnant uterus, suggesting the effect on decidualization was independent of the conceptus. This included a suppression of endometrial stroma cell proliferation and of key decidualization regulators, including COX2, BMP2 and WNT4. Collectively, these data demonstrate that maternal epinephrine exposure during early pregnancy impairs uterine decidualization and embryo development, underlying early pregnancy failure.


Pharmacologic treatment of neuropathic pain.
Neuropathic pain, or pain after nervous system injury, can be very refractory to pharmacologic interventions. Through a better understanding of the pathophysiology of neuropathic pain, it has been suggested that nonopioid agents, such as antidepressants and anticonvulsants, may be more efficacious in the treatment of neuropathic pain than common analgesics, such as opioids or nonsteroidal anti-inflammatory drugs. However, this has not been consistently demonstrated in clinical studies. Conversely, many confounding factors of neuropathic pain make it difficult to interpret clinical studies. Therefore, we must develop a better understanding of the preclinical models of neuropathic pain to better understand the application of new and old drugs to the human neuropathic pain state. This article provides an overview of the commonly used preclinical neuropathic pain models, followed by a summary of the efficacy of currently available agents in preclinical pain models and human correlates.


The importance of a medical chaperone: a quality improvement study exploring the use of a note stamp in a tertiary breast surgery unit.
The project aim was to determine current use and documentation of medical chaperones within a major breast service unit. It explored ways of improving adherence to professional guidelines concerning chaperones. The single centre quality improvement project was completed in a tertiary breast service unit in North West London. It was a three-stage project with initial audit in October 2013, 1st postintervention cycle in November 2013 and 2nd postintervention cycle in October 2014. In each study cycle, data were collected from entries in clinic notes until at least 155 encounters with documented clinical examination were analysed. All notes were of female patients. (1) Intervention 1st cycle: presentation and discussion of chaperone guidelines alongside reminder posters and introduction of note stamp. (2) Intervention 2nd cycle: note stamp alone. Documentation of chaperone offer, documentation of patient preference regarding chaperone, identifier (name or signature) of chaperone present and gender of examining clinician. In the 1st postintervention cycle, 69.95% documentation of chaperone offer was recorded, p<0.001, CI (59.04% to 80.76%). This result was replicated in the 2nd postintervention cycle a year later with 74.86% documentation of chaperone offer recorded, p<0.001, CI (66.41% to 83.31%). The 4.91% difference was insignificant; p=0.294, CI (14.03% to 4.21%). The authors suggest that a proforma approach to medical chaperones is an effective means of ensuring adherence to best practice guidelines. A stamp, or similar, that can be embedded into documentation structure is an effective example of such an approach. Improved documentation allows any problems with adherence to guidelines to be more easily identified, helping to ensure the safeguarding of patients and staff involved in intimate examinations.


Genetic testing practices in infants with congenital heart disease.
Clinical genetic testing is expanding rapidly, but the application of new testing has not been reported in an unselected, comprehensive congenital heart disease (CHD) patient population. This study aims to identify cytogenetic testing practices and diagnostic yield in infants with CHD as an important first step toward understanding clinical utility of dedicated cytogenetic testing. We hypothesized that chromosome microarray analysis (CMA) would identify genetic abnormalities underlying both syndromic and isolated CHD. This is a single institution retrospective study that characterizes cytogenetic testing practices and diagnostic yield for all cytogenetic testing in each infant identified with CHD over a 32-month period. CHD was classified by type, complexity, and presence or absence of extracardiac anomalies. Among the 1087 infants identified with CHD by echocardiogram, 277 infants (25%) had some form of cytogenetic testing, including karyotype, fluorescence in situ hybridization, and/or CMA. Forty-one percent of infants who had cytogenetic testing had more than one test. CMA was performed in 121 patients (11%), and abnormalities (both clinically significant and variants of unknown significance) were identified in 35/121 (29%). Forty-nine percent of CMA abnormalities were in patients with apparently isolated nonsyndromic CHD. This single institution study identified that only 25% of infants with CHD underwent cytogenetic testing, indicating possible underutilization of testing in this age group. The high multiple testing rate indicates a need for improved guidelines for cost effective testing approaches. The diagnostic yield in this study suggests that CMA is a particularly useful first screening test when a specific syndrome is not clinically identifiable. Larger studies investigating cardiac lesion-specific diagnostic yield in isolated CHD are warranted.


Splicing of retrotransposon insertions from transcripts of the Drosophila melanogaster vermilion gene in a revertant.
A mutation of the Drosophila melanogaster vermilion (v) gene known as v1 is caused by the insertion of a 412 retrotransposon into the 5' untranslated region of the first exon. Mutants carrying this insertion accumulate a low level of mRNA from which most of the transposon sequences have been eliminated by splicing at cryptic sites within transposon sequences. Here, we demonstrate that a revertant of the v1 allele called v+37 is caused by the insertion of a second retrotransposon, the B104/roo element, into a site near one end of the 412 element. The revertant strain accumulates a higher level of mRNA from which most of both transposons have been removed by splicing at new donor sites introduced by the B104/roo insertion and the same acceptor site within 412. Mutations at suppressor of sable [su(s)], which increase the accumulation of v1 transcripts, slightly elevate the level of v+37 RNA. In addition, we show that the first v intron downstream of the 412 insertion is not efficiently removed in the v1 mutant, and suppressor and reversion mutations increase the proportion of transcripts that are properly spliced at that downstream intron. Thus, it appears that both the suppressor and reversion mutations exert an effect at the level of pre-mRNA splicing.


[Motor asymmetry and hemisphere interactions in Parkinson's disease].
About 90% of the people in general population are right-handed. Such right-side motor lateralization is presumed to be, at least partially, related to higher brain functions, especially to presentation of speech areas in the left hemisphere. Although lateralization may be predetermined genetically, the investigations into this problem have not yielded the equivocal results. Motor deficit asymmetry is a constant trait of Parkinson's disease (PD). It means that a study of PD patients gives a remarkable possibility for investigation into hemisphere interactions. Using motor potential method, 62 patients with predominantly akinetico-rigid lateralized PD type and relatively preserved social adaptation have been examined. The patients differed from tho controls by all MP components and reduction of hemisphere asymmetry: while using both right and left hands counterlateral hemisphere, as well as ipsilateral one, were insufficiently activated. This indicates pathological process diffusion in PD and hemisphere interaction dysfunction. Even in hemiparkinsonian stage of PD, a rather unusual pattern of hemisphere interaction is formed.


Diminishing Clinical Returns of Multilevel Minimally Invasive Lumbar Interbody Fusion.
Single institution retrospective clinical review. To investigate the relationship between levels fused and clinical outcomes in patients undergoing open and minimally invasive surgical (MIS) lumbar fusion. Minimally invasive spinal fusion aims to reduce the morbidity associated with conventional open surgery. As multilevel arthrodesis procedures are increasingly performed using MIS techniques, it is necessary to weigh the risks and benefits of multilevel MIS lumbar fusion as a function of fusion length. Patients undergoing <4 level lumbar interbody fusion were stratified by surgical technique (MIS or open), and grouped by fusion length: 1-level, 2-levels, 3+ levels. Demographics, Charlson Comorbidity Index (CCI), surgical factors, and perioperative complication rates were compared between technique groups at different fusion lengths using means comparison tests. Included: 361 patients undergoing lumbar interbody fusion (88% transforaminal, 14% lateral; 41% MIS). Breakdown by fusion length: 63% 1-level, 22% 2-level, 15% 3+ level. Op-time did not differ between groups at 1-level (MIS: 233 min vs. Open: 227, P = 0.554), though MIS at 2-levels (332 min vs. 281) and 3+ levels (373 min vs. 323) were longer (P = 0.033 and P = 0.231, respectively). While complication rates were lower for MIS at 1-level (15% vs. 30%, P = 0.006) and 2-levels (13% vs. 27%, P = 0.147), at 3+ levels, complication rates were comparable (38% vs. 35%, P = 0.870). 3+ level MIS fusions had higher rates of ileus (13% vs. 0%, P = 0.008) and a trend of increased adverse pulmonary events (25% vs. 7%, P = 0.110). MIS was associated with less EBL at all lengths (all P < 0.01) and lower rates of anemia at 1-level (5% vs. 18%, P < 0.001) and 2-levels (7% vs. 16%, P = 0.193). At 3+ levels, however, anemia rates were similar between groups (13% vs. 15%, P = 0.877). MIS lumbar interbody fusions provided diminishing clinical returns for multilevel procedures. While MIS patients had lower rates of perioperative complications for 1- and 2-level fusions, 3+ level MIS fusions had comparable complication rates to open cases, and higher rates of adverse pulmonary and ileus events. 3.


Predictive value of the transtheoretical model to smoking cessation in hospitalized patients with cardiovascular disease.
Several authors have questioned the transtheoretical model. Determining the predictive value of each cognitive-behavioural element within this model could explain the multiple successes reported in smoking cessation programmes. The purpose of this study was to predict point-prevalent smoking abstinence at 2 and 6 months, using the constructs of the transtheoretical model, when applied to a pooled sample of individuals who were hospitalized for a cardiovascular event. The study follows a predictive correlation design. Recently hospitalized patients (n=168) with cardiovascular disease were pooled from a randomized, controlled trial. Independent variables of the predictive transtheoretical model comprise stages and processes of change, pros and cons to quit smoking (decisional balance), self-efficacy, and social support. These were evaluated at baseline, 2 and 6 months. Compared to smokers, individuals who abstained from smoking at 2 and 6 months were more confident at baseline to remain non-smokers, perceived less pros and cons to continue smoking, utilized less consciousness raising and self-re-evaluation experiential processes of change, and received more positive reinforcement from their social network with regard to their smoke-free behaviour. Self-efficacy and stages of change at baseline were predictive of smoking abstinence after 6 months. Other variables found to be predictive of smoking abstinence at 6 months were an increase in self-efficacy; an increase in positive social support behaviour and a decrease of the pros within the decisional balance. The results partially support the predictive value of the transtheoretical model constructs in smoking cessation for cardiovascular disease patients.


Next-generation flexible formats of VNAR domains expand the drug platform's utility and developability.
Therapeutic mAbs have delivered several blockbuster drugs in oncology and autoimmune inflammatory disease. Revenue for mAbs continues to rise, even in the face of competition from a growing portfolio of biosimilars. Despite this success, there are still limitations associated with the use of mAbs as therapeutic molecules. With a molecular mass of 150 kDa, a two-chain structure and complex glycosylation these challenges include a high cost of goods, limited delivery options, and poor solid tumour penetration. There remains an urgency to create alternatives to antibody scaffolds in a bid to circumvent these limitations, while maintaining or improving the therapeutic success of conventional mAb formats. Smaller, less complex binders, with increased domain valency, multi-specific/paratopic targeting, tuneable serum half-life and low inherent immunogenicity are a few of the characteristics being explored by the next generation of biologic molecules. One novel 'antibody-like' binder that has naturally evolved over 450 million years is the variable new antigen receptor (VNAR) identified as a key component of the adaptive immune system of sharks. At only 11 kDa, these single-domain structures are the smallest IgG-like proteins in the animal kingdom and provide an excellent platform for molecular engineering and biologics drug discovery. VNAR attributes include high affinity for target, ease of expression, stability, solubility, multi-specificity, and increased potential for solid tissue penetration. This review article documents the recent drug developmental milestones achieved for therapeutic VNARs and highlights the first reported evidence of the efficacy of these domains in clinically relevant models of disease.


The law and politics of embryo research in America.
The moral, legal, and public policy dispute over embryonic stem cell research (and related matters, such as human cloning) is the most prominent issue in American public bioethics of the past decade. The primary moral question raised by the practice of embryonic stem cell research is whether it is defensible to disaggregate (and thus destroy) living human embryos in order to derive pluripotent cells (stem cells) for purposes of basic research that may someday yield regenerative therapies. This essay will explain the legal and political dimensions of the embryonic stem cell debate as it has unfolded at the national level in the United States, contrasting the position and thinking of President Clinton's administration with that of George W Bush. Building upon this, a set of brief reflections is offered on the form and substance of the American federal approach to this public matter and whether it is ultimately sustainable to join the issue in this particular way.


Effects of vaccine route and dosage on protection from rabies after intracerebral challenge in mice.
To evaluate the effect of various routes of administration and number of doses of 3 commercially produced rabies vaccines on serum antibody responses and protection in mice challenged by intracerebral injection with fixed-strain rabies virus. 2,213 mice. Inactivated, adjuvanted rabies vaccines were administered to mice in either 2, 1, or 0 (control) doses via IP, IM, and SC routes, and mice were challenged intracerebrally with fixed-strain rabies virus. Vaccination route and dose number significantly influenced serum antibody responses and protection from rabies virus challenge, independent of vaccine strain origin and mouse strain, although mouse age significantly affected results. Extended challenge studies revealed that IM vaccination of mice resulted in the highest serum neutralizing antibody responses and protection levels equivalent to IP vaccination. Even multiple doses administered SC (a vaccination route used in dogs) resulted in poor serum anti-rabies neutralizing antibody responses in mice and were far less protective than other routes. Findings suggest possible improvements for the current rabies vaccine potency test in mice by using 1 dose, the IM route, and a delayed time of challenge. These modifications would more closely model vaccination practices in target species and yield more accurate information regarding primary immunogenicity of a vaccine.


Hydroxysafflor yellow a attenuates small airway remodeling in a rat model of chronic obstructive pulmonary disease.
Our previous studies found that hydroxysafflor yellow A (HSYA), an active ingredient in Carthamus tinctorius L., has anti-inflammatory and anti-fibrosis properties. In this study, we investigated the effect of HSYA on small airway remodeling (SAR) in a chronic obstructive pulmonary disease (COPD) rat model induced by cigarette smoke and lipopolysaccharide (LPS). SAR is a common lesion in COPD characterized by thickening of the airway wall, mainly by subepithelial fibrosis. In this study the thickness of the small airway was determined by total wall area/basement membrane perimeter (WAt/Pbm). Collagen deposition of the small airway was assessed by Masson's trichrome staining. HSYA significantly attenuated the thickening and collagen deposition of the small airway and inhibited transforming growth factor β1 (TGF-β1) mRNA and protein expression in COPD rat. In addition, HSYA inhibited the phosphorylation of p38 mitogen-activated protein kinases (MAPK) in the lung tissue of rat. HSYA can attenuate experimentally induced airway remodeling and this attenuation may be attributed to suppression of TGF-β1 expression.


Views of primary care physicians and oncologists on cancer follow-up initiatives in primary care: an online survey.
Primary care physicians (PCPs) are playing an increasing role in the follow-up of cancer in England. In 2003 a Cancer Care Review (CCR) was introduced to ensure contact between PCPs and cancer patients within 6 months of diagnosis. The NHS also intends to introduce survivorship care plans (SCP). The aims of this study were to: describe current practice and views in primary care with respect to the CCR and information provision from secondary to primary care following final discharge from hospital follow-up; and to seek views on the perceived usefulness, content, and feasibility of a SCP. An on-line questionnaire survey of 100 oncologists and 200 PCPs. Half of PCPs undertook the CCR opportunistically, and only 64% had an agreed structure. Forty percent felt the CCR was useful for the doctor, and 60% useful for the patient. Most PCPs and oncologists think a SCP would be useful, but only 40% oncologists thought that it would be easy to produce. At discharge from follow-up, more than half of oncologists said they provided information on histology, treatment, requirements for screening and surveillance, and referral guidance. Less than half provide information on potential late effects and symptoms of recurrence. PCPs felt that information on all of these areas was important and that the information they receive is often inadequate. The CCR has not been implemented systematically. There is support for the introduction of a SCP and broad agreement on content. However, careful planning is needed to ensure all necessary information is included and to overcome barriers of implementation. Further research should explore what cancer survivors would find useful in a primary care-based CCR and what should be included in a SCP. This should be clearly communicated to the relevant health care professionals to maximise the benefits cancer survivors and their families gain from these policy initiatives.


Novel endogenous negative modulators of platelet function as potential anti-thrombotic targets.
Platelets are megakaryocyte-derived nuclear-free fragments that participate in cardiovascular diseases including acute myocardial infarction, ischemic stroke, hypertension, and atherosclerosis. At the endothelium damage site, platelets interact with sub-endothelial matrix proteins such as glycoprotein VI/Fc receptor γ-chain (GPVI/FcRγ), G protein-coupled receptor/phospholipase Cγ(β) (GPCR/PLCγ(β)), Rho/RhoK and integrin. The activation of these signaling pathways triggers intracellular calcium increase and causes platelet adhesion, aggregation, granule release and finally thrombus formation. Some endogenous platelet modulators reported to negatively regulate this process are: (1) platelet surface inhibitory receptors: carcinoembryonic antigen cell adhesion molecule 1, 2 (CEACAM 1, 2), platelet endothelial cell adhesion molecule-1 (PECAM-1), and G6b-B; (2) nuclear receptors: retinoic X receptor (RXRs), glucocorticoid receptor (GR), peroxisome proliferator-activated receptors (PPARs) and liver X receptors (LXRs); (3) intracellular adaptor proteins: CLP36, paxillin, downstream of tyrosine kinase (Dok), c-Casitas B-lineage lymphoma (c-Cbl), protein kinase Cδ (PKCδ), glycogen synthase kinase(GSK)-3β, phospholipase D2 (PLD2), peroxiredoxin II (PrxII), T-cell ubiquitin ligand-2 (TULA-2); (4) extracellular modulators released from platelet granules: adapter protein disabled-2 (DAB2) and diadenosine 5,5-P1, P2-diphosphate (Ap2A). The discovery of biological or endogenous modulators of platelet activation is regarded as a potential therapeutic target for thrombotic disease. This review highlights the recent findings on the endogenous negative regulatory molecules released from platelets and their impact on platelet thrombus formation.


Atomic-Scale Modulation of Synthetic Magnetic Order in Oxide Superlattices.
Atomic-scale precision control of magnetic interactions facilitates a synthetic spin order useful for spintronics, including advanced memory and quantum logic devices. Conventional modulation of synthetic spin order has been limited to metallic heterostructures that exploit Ruderman-Kittel-Kasuya-Yosida interaction through a nonmagnetic metallic spacer; however, they face issues arising from Joule heating and/or electric breakdown. The practical realization and observation of a synthetic spin order across a nonmagnetic insulating spacer will lead to the development of spin-related devices with a completely different concept. Herein, the atomic-scale modulation of the synthetic spiral spin order in oxide superlattices composed of ferromagnetic metal and nonmagnetic insulator layers is reported. The atomically controlled superlattice exhibits an oscillatory magnetic behavior, representing the existence of a spiral spin structure. Depth-sensitive polarized neutron reflectometry evidences modulated spiral spin structures as a function of the nonmagnetic insulator layer thickness. Atomic-scale customization of the spin state can move the field one step further to actual spintronic applications.


Polyomavirus infection in a free-ranging California sea lion (Zalophus californianus) with intestinal T-cell lymphoma.
An adult female California sea lion (Zalophus californianus) that stranded in central California was found to have a small glossal polypoid mass on gross necropsy. Histologically, the mass was consistent with a fibropapilloma, and intranuclear inclusions were found within endothelial cells lining small arterioles within the mass. Electron microscopy revealed 40-nm virions within endothelial intranuclear inclusions. Rolling circle amplification was used to obtain a partial viral genomic sequence. Sequence analysis identified the virus as a novel polyomavirus, tentatively named California sea lion polyomavirus 1. In addition, the sea lion had a severely thickened small intestine and swollen pale kidneys on gross examination. Severe renal amyloidosis with chronic interstitial nephritis was diagnosed histologically as well as T-cell intestinal lymphoma, which was confirmed via immunophenotyping and molecular clonality. The relationship, if any, between polyomavirus infection and the other disease processes in this sea lion is not known, but it is considered unlikely that the polyomavirus induced the lymphoma.


LPS Stimulation of Cord Blood Reveals a Newborn-Specific Neutrophil Transcriptomic Response and Cytokine Production.
The neonatal innate immune system differs to microbial infection both quantitatively and qualitatively when compared with adults. Here, we provide the first genome-wide ex-vivo expression profile of umbilical cord blood (UCB) neutrophils from full-term infants prior to and in response to whole-blood lipopolysaccharide (LPS) stimulation. Additionally, we provide cytokine expression prior to and following LPS stimulation. The genomic expression and cytokine profile are compared with LPS-stimulated whole blood from healthy adult subjects (HC). Whole blood from UCB (n = 6) and HC (n = 6) was studied at baseline or was stimulated for 24 h with 100 ngs/mL of LPS. CD66b neutrophils were subsequently isolated with microfluidic techniques and genome-wide expression analyses were performed. Ingenuity Pathway Analysis (IPA) software was utilized to predict downstream functional effects. Additionally, cytokine concentrations in whole blood prior to and after 24 h of LPS incubation were determined. LPS stimulated whole blood from UCB demonstrated significant differences in both ex-vivo cytokine production and PMN gene expression. Mixed-effect modeling identified 1,153 genes whose expression changed significantly in UCB and HC after exposure to LPS (P < 0.001 with a minimum 1.5-fold change). IPA downstream predictions suggest that PMNs from UCB fail to effectively upregulate genes associated with activation, phagocytosis, and chemotaxis in response to LPS stimulation. Furthermore, whole blood from UCB showed increased interleukin (IL)-10 production to LPS, but failed to significantly increase several pro-inflammatory cytokines. LPS-stimulated whole blood from UCB exhibited a markedly suppressed inflammatory cytokine production and PMN innate immune genome response. These differences in gene expression and cytokine production may be an adaptive response to a prior fetal environment, but may also explain their increased susceptibility to infections. Characterization of these deficits is the first step toward developing prophylactic and therapeutic interventions.


The light and electron microscopic localization of intracytoplasmic nucleolus-like bodies in the mouse brain stained by Holmes' silver method.
By light microscopic observation of the mouse brain impregnated by the Holmes' method, intensely stained cytoplasmic inclusion bodies were demonstrated in the nerve cells of the hypothalamus, locus coeruleus and other autonomic regions in addition to the limbic system and some circumventricular organs. Electron microscope observation of ultrathin sections made from vibratome sections impregnated by the Holmes' method revealed that the structures which have been called the nucleolus-like bodies were loaded with large amounts of gold particles. Hence, the Holmes-impregnated cytoplasmic inclusions could be identified with the so-called nucleolus-like bodies. The fine structure, distribution and significance of these bodies were described.


Evidence for presence of functional beta-adrenoceptor in rabbit S2 proximal straight tubules.
The effect of isoproterenol on the electrophysiological properties of the S2 proximal segment of the rabbit was examined. Isoproterenol at 10(-8) to 10(-4) M depolarized the basolateral membrane voltage (Vb) in a dose-dependent manner. Propranolol attenuated the isoproterenol-induced depolarization. These possible mechanisms of cell depolarization were explored. The role of luminal Na(+)-organic solute cotransport was negligible, since the removal of organic solute did not change the depolarization. Basolateral Na(+)-(HCO3-) cotransport was supported by the finding that 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid inhibited isoproterenol-induced depolarization. Basolateral K+ conductance was suggested by the finding that the application of Ba2+ blocked the isoproterenol-induced depolarization. Na(+)-K(+)-adenosine-triphosphatase (ATPase) was questionable. Although ouabain blocked isoproterenol-induced depolarization, the removal of Na+ did not inhibit the depolarization. Further experiment revealed that dibutylyl-adenosine 3',5'-cyclic monophosphate (cAMP), 8-bromo cAMP, and forskolin did not mimic the response of isoproterenol. These results demonstrate: 1) there is a functional beta-adrenoceptor that depolarizes Vb; 2) isoproterenol-induced depolarization is due to an inhibition of basolateral K+ channel or the activation of basolateral Na(+)-(HCO3-)n cotransport; 3) isoproteronol-induced depolarization is independent of cAMP in the rabbit proximal tubule.


Monolithic integration of III-V nanowire with photonic crystal microcavity for vertical light emission.
A novel photonic structure formed by the monolithic integration of a vertical III-V nanowire on top of a L3 two-dimensional photonic crystal microcavity is proposed to enhance light emission from the nanowire. The impact on the nanowire spontaneous emission rate is evaluated by calculating the spontaneous emission factor β, and the material gain at threshold is used as a figure of merit of this vertical emitting nanolaser. An optimal design is identified for a GaAs nanowire geometry with r = 155 nm and L~1.1 μm, where minimum gain at threshold (gth~13×10³ cm⁻¹) and large spontaneous emission factor (β~0.3) are simultaneously achieved. Modification of the directivity of the L3 photonic crystal cavity via the band-folding principle is employed to further optimize the far-field radiation pattern and to increase the directivity of the device. These results lay the foundation for a new approach toward large-scale integration of vertical emitting nanolasers and may enable applications such as intra-chip optical interconnects.


Strength training improves upper-limb function in individuals with stroke: a meta-analysis.
After stroke, maximal voluntary force is reduced in the arm and hand muscles, and upper-limb strength training is 1 intervention with the potential to improve function. We performed a meta-analysis of randomized controlled trials. Electronic databases were searched from 1950 through April 2009. Strength training articles were assessed according to outcomes: strength, upper-limb function, and activities of daily living. The standardized mean difference (SMD) was calculated to estimate the pooled effect size with random-effect models. From the 650 trials identified, 13 were included in this review, totaling 517 individuals. A positive outcome for strength training was found for grip strength (SMD=0.95, P=0.04) and upper-limb function (SMD=0.21, P=0.03). No treatment effect was found for strength training on measures of activities of daily living. A significant effect for strength training on upper-limb function was found for studies including subjects with moderate (SMD=0.45, P=0.03) and mild (SMD=0.26, P=0.01) upper-limb motor impairment. No trials reported adverse effects. There is evidence that strength training can improve upper-limb strength and function without increasing tone or pain in individuals with stroke.


Prediction of probabilistic sleep distributions following travel across multiple time zones.
To parameterize and validate a model to estimate average sleep times for long-haul aviation pilots during layovers following travel across multiple time zones. The model equations were based on a weighted distribution of domicile- and local-time sleepers, and included algorithms to account for sleep loss and circadian re-synchronization. Sleep times were collected from participants under normal commercial operating conditions using diaries and wrist activity monitors. Participants included a total of 306 long-haul pilots (113 captains, 120 first officers, and 73 second officers). The model was parameterized based on the average sleep/wake times observed during international flight patterns from Australia to London and Los Angeles (global R2 = 0.72). The parameterized model was validated against the average sleep/wake times observed during flight patterns from Australia to London (r2 = 0.85), Los Angeles (r2 = 0.79), New York (r2 = 0.80), and Johannesburg (r2 = 0.73). Goodness-of-fit was poorer when the parameterized model equations were used to predict the variance across the sleep/wake cycles of individual pilots (R2 = 0.42, 0.35, 0.31, and 0.28 for the validation flight patterns, respectively), in part because of substantial inter-individual variability in sleep timing and duration. It is possible to estimate average sleep times during layovers in international patterns with a reasonable degree of accuracy. Models of this type could form the basis of a stand-alone application to estimate the likelihood that a given duty schedule provides pilots, on average, with an adequate opportunity to sleep.


Slicing Through the Layers of Pediatric Unintentional Injury With the Swiss Cheese Model: A Topical Review.
Pediatric unintentional injury significantly burdens children, families, and society. Behavioral researchers have examined the problem for decades, identifying many risk factors and greatly enhancing knowledge of the injury antecedent process. Approaches using theories and models to guide inquiry into etiology and prevention are still needed. We offer an approach borrowed from the field of human factors to enhance understanding and prevention. We focus our exploration on an error modeling and accident investigation tool called the Swiss Cheese Model. We first introduce the basic elements of the model. Next, we apply error modeling concepts to example scenarios drawn from real unintentional incidents and discuss the implications for understanding etiology and prevention. Finally, potential future directions are discussed to illustrate paths for the advancement of injury etiology and prevention.


A study and optimization of lumbar spine X-ray imaging systems.
A Monte Carlo program has been developed that incorporates a voxel phantom of an adult patient in a model of the complete X-ray imaging system, including the anti-scatter grid and screen-film receptor. This allows the realistic estimation of patient dose and the corresponding image (optical density map) for a wide range of equipment configurations. This paper focuses on the application of the program to lumbar spine anteroposterior and lateral screen-film examinations. The program has been applied to study the variation of physical image quality measures and effective dose for changing system parameters such as tube voltage, grid design and screen-film system speed. These variations form the basis for optimization of these system parameters. In our approach to optimization, the best systems are those that can match (or come close to) the calculated image quality measure of systems preferred in a recent European clinical trial, but with lower patient dose. The largest dose savings found were 21% for a 400 speed class system with a grid having a strip density of 40 cm(-1) and a grid ratio of 16. A further dose saving of 13% was possible when a 600 speed class system was employed. The best systems found from the optimization correspond to those recommended by the European Commission guidelines on image quality criteria for diagnostic radiographic images.


Acute Pancreatitis: A Review.
In the United States, acute pancreatitis is one of the leading causes of hospital admission from gastrointestinal diseases, with approximately 300 000 emergency department visits each year. Outcomes from acute pancreatitis are influenced by risk stratification, fluid and nutritional management, and follow-up care and risk-reduction strategies, which are the subject of this review. MEDLINE was searched via PubMed as was the Cochrane databases for English-language studies published between January 2009 and August 2020 for current recommendations for predictive scoring tools, fluid management and nutrition, and follow-up and risk-reduction strategies for acute pancreatitis. Several scoring systems, such as the Bedside Index of Severity in Acute Pancreatitis (BISAP) and the Acute Physiology and Chronic Health Evaluation (APACHE) II tools, have good predictive capabilities for disease severity (mild, moderately severe, and severe per the revised Atlanta classification) and mortality, but no one tool works well for all forms of acute pancreatitis. Early and aggressive fluid resuscitation and early enteral nutrition are associated with lower rates of mortality and infectious complications, yet the optimal type and rate of fluid resuscitation have yet to be determined. The underlying etiology of acute pancreatitis should be sought in all patients, and risk-reduction strategies, such as cholecystectomy and alcohol cessation counseling, should be used during and after hospitalization for acute pancreatitis. Acute pancreatitis is a complex disease that varies in severity and course. Prompt diagnosis and stratification of severity influence proper management. Scoring systems are useful adjuncts but should not supersede clinical judgment. Fluid management and nutrition are very important aspects of care for acute pancreatitis.


[Methods of hormone replacement therapy in hypertensive women].
A comparative retrospective study was made to examine influence of replacement therapy by estradiol valeriate and climonorm on perimenopausal hypertension. The test group (25 hypertensive women) received climonorm for 6 months. Control women received hypotensive therapy. Lipid metabolism and risk of vascular events were assessed. The replacement therapy had a positive effect on lipid metabolism. Criteria of Mann-Whitney, Friedmann, Wilkinson and of chi 2 were made use of. The target arterial pressure was achieved only with hypotensive therapy which also diminished intensity of menopausal disorders. To determine individual policy, it is necessary to stratify risk of cardiovascular events. The best effect is achieved at combination of hypotensive therapy with replacement hormone treatment.


Effects of a naturally occurring mutation in the hepatitis B virus basal core promoter on precore gene expression and viral replication.
The basal core promoter (BCP) of hepatitis B virus (HBV) controls the transcription of both the precore RNA and the core RNA. The precore RNA codes for the secreted e antigen, while the core RNA codes for the major core protein and the DNA polymerase and also is the pregenomic RNA. The double mutation of nucleotides 1762 and 1764 in the BCP from A and G to T and A, respectively, is frequently observed in HBV sequences isolated from chronic patients. Several papers have reported conflicting results regarding whether this double mutation is important for e antigen expression. In order to address this issue, we have introduced this double mutation into the HBV genome and studied its effects on HBV gene expression and replication. Our results indicate that the mutated BCP can no longer bind a liver-enriched transcription factor(s) and that the transcription of only precore RNA and, consequently, the expression of e antigen were reduced. The reduction of precore gene expression was accompanied by an increase in progeny virus production. This increase was found to occur at or immediately prior to the encapsidation of the pregenomic RNA. Thus, the results of our in vitro study resolve the discrepancy of previous clinical observations and indicate that this double mutation suppresses but does not abolish the e antigen phenotype. The implications of these findings in the pathogenesis of HBV are discussed.


Observation of an Excess of Dicharmonium Events in the Four-Muon Final State with the ATLAS Detector.
A search is made for potential ccc[over ¯]c[over ¯] tetraquarks decaying into a pair of charmonium states in the four muon final state using proton-proton collision data at sqrt[s]=13 TeV, corresponding to an integrated luminosity of 140 fb^{-1} recorded by the ATLAS experiment at LHC. Two decay channels, J/ψ+J/ψ→4μ and J/ψ+ψ(2S)→4μ, are studied. Backgrounds are estimated based on a hybrid approach involving Monte Carlo simulations and data-driven methods. Statistically significant excesses with respect to backgrounds dominated by the single parton scattering are seen in the di-J/ψ channel consistent with a narrow resonance at 6.9 GeV and a broader structure at lower mass. A statistically significant excess is also seen in the J/ψ+ψ(2S) channel. The fitted masses and decay widths of the structures are reported.


Oestradiol inhibits collagen breakdown in the involuting rat uterus.
1. The earlier observation (Woessner, 1969) of oestradiol inhibition of collagen breakdown is confirmed and extended. Administration of 100mug of oestradiol-17beta/day to parturient rats strongly inhibits the loss of collagen from the involuting uterus. Three experiments show that this effect is due to an inhibition of collagen degradation rather than to a stimulation of collagen synthesis. 2. Uterine collagen was labelled with hydroxy[(14)C]-proline by the administration of [(14)C]proline near the end of pregnancy. By 3 days post partum, control uteri lost 83% of their collagen and 90% of their hydroxy[(14)C]proline. Uteri from oestradiol-treated rats lost only 50% of both total and labelled hydroxyproline, with no decrease in the specific radioactivity of the hydroxyproline. 3. Incorporation of [(14)C]proline into uterine collagen hydroxyproline in vivo was not affected by oestradiol treatment. 4. Urinary excretion of hydroxyproline was increased in post-partum control rats and decreased in oestradiol-treated rats. 5. An enzyme capable of cleaving 4-phenylazobenzyloxycarbonyl-l-prolyl-l-leucylglycyl- l-prolyl-d-arginine (a substrate for clostridial collagenase) increased in activity in the post-partum uterus and was unaffected by oestradiol treatment. 6. Uterine homogenates digested uterine collagen extensively at pH3.2. This digestion was unaffected by the oestradiol treatment. 7. Lysosomal fractions prepared by density-gradient centrifugation of uterine homogenates contained coincident peaks of cathepsin D activity and peptide-bound hydroxyproline. The cathepsin D and hydroxyproline contents of this peak were unaffected by oestradiol treatment.


Interactions between protein kinase CK2 and Pin1. Evidence for phosphorylation-dependent interactions.
The peptidyl-prolyl isomerase Pin1 interacts in a phosphorylation-dependent manner with several proteins involved in cell cycle events. In this study, we demonstrate that Pin1 interacts with protein kinase CK2, an enzyme that generally exists in tetrameric complexes composed of two catalytic CK2 alpha and/or CK2 alpha' subunits together with two regulatory CK2 beta subunits. Our results indicate that Pin1 can interact with CK2 complexes that contain CK2 alpha. Furthermore, Pin1 can interact directly with the C-terminal domain of CK2 alpha that contains residues that are phosphorylated in vitro by p34(Cdc2) and in mitotic cells. Substitution of the phosphorylation sites of CK2 alpha with alanines resulted in decreased interactions between Pin1 and CK2. The other catalytic isoform of CK2, designated CK2 alpha', is not phosphorylated in mitotic cells and does not interact with Pin1, but a chimeric protein consisting of CK2 alpha' with the C terminus of CK2 alpha was phosphorylated in mitotic cells and interacts with Pin1, further implicating the phosphorylation sites in the interaction. In vitro, Pin1 inhibits the phosphorylation of Thr-1342 on human topoisomerase II alpha by CK2. Topoisomerase II alpha also interacts with Pin1 suggesting that the effect of Pin1 on the phosphorylation of Thr-1342 could result from its interactions with CK2 and/or topoisomerase II alpha. As compared with wild-type Pin1, isomerase-deficient and WW domain-deficient mutants of Pin1 are impaired in their ability to interact with CK2 and to inhibit the CK2-catalyzed phosphorylation of topoisomerase II alpha. Collectively, these results indicate that Pin1 and CK2 alpha interact and suggest a possible role for Pin1 in the regulation of topoisomerase II alpha. Furthermore, these results provide new insights into the functional role of the mitotic phosphorylation of CK2 and provide a new mechanism for selectively regulating the ability of CK2 to phosphorylate one of its mitotic targets.


Alveolar macrophage and glycoprotein responses to Pneumocystis carinii.
Pneumocystis carinii continues to represent an important complication of immunosuppression in patients with acquired immune deficiency syndrome, hematological and solid malignancies, organ transplantation, and during corticosteroid and cytotoxic therapy for inflammatory disorders. Although host defenses against this organism center around CD4 lymphocytic function, additional immune mediators in the alveolar spaces contribute substantially to host recognition and elimination of P. carinii. In particular, this review considers the interactions of P. carinii with alveolar macrophages, adhesive glycoproteins including vitronectin and fibronectin, and surfactant lipids and protein components. Recent studies indicate that alveolar macrophages contribute significantly to host responses against this organism by mediating uptake and degradation of P. carinii, and by releasing inflammatory mediators including reactive oxidants, eicosanoids, and the potent proinflammatory cytokine tumor necrosis factor-alpha. Furthermore, the interactions of P. carinii with multiple adhesive proteins and with surfactant components additionally modulate the interactions of P. carinii with macrophages and enhance host recognition of this pathogen. These non-lymphocytic mediators represent additional important mechanisms of host recognition and response to P. carinii infection.


Pyoderma gangrenosum.
A 3.5-year-old girl presented with skin lesions beginning as a small pustule that progressed to very painful large ulcer with rolled up edges and irregular borders. At presentation, active ulcers were present over the left elbow. There were healed ulcers with scarring present on the chest, back, both thighs and abdominal wall. This child had negative antinuclear antibody, cytoplasmic antinuclear cytoplasmic antibody, antiphospholipid antibodies and tests for hepatitis B and C. She did not have any symptoms to suggest ulcerative colitis. Her colonoscopy was absolutely normal. Histopathology from her skin lesion was consistent with pyoderma gangrenosum.


Telematic integration of health data: a practicable contribution.
The patients' clinical and healthcare data should virtually be available everywhere, both to provide a more efficient and effective medical approach to their pathologies, as well as to make public healthcare decision makers able to verify the efficacy and efficiency of the adopted healthcare processes. Unfortunately, customised solutions adopted by many local Health Information Systems in Italy make it difficult to share the stored data outside their own environment. In the last years, worldwide initiatives have aimed to overcome such sharing limitation. An important issue during the passage towards standardised, integrated information systems is the possible loss of previously collected data. The herein presented project realises a suitable architecture able to guarantee reliable, automatic, user-transparent storing and retrieval of information from both modern and legacy systems. The technical and management solutions provided by the project avoid data loss and overlapping, and allow data integration and organisation suitable for data-mining and data-warehousing analysis.


Transient ventilatory and heart rate responses to moderate nonabrupt pseudorandom exercise.
Dynamic responses of inspired minute ventilation, CO2 and O2 end-tidal gas fractions, and heart rate were obtained from six normal human volunteers in response to a complex dynamic exercise challenge. Subjects pedalled a chair ergometer at constant frequency. The retarding torque applied to the ergometer pedals was controlled by a low-pass-filtered pseudorandom binary sequence (fPRBS), which provided a complex, nonanticipatory exercise stimulus containing sufficient high- and low-frequency energy to excite the small signal, broadband ventilatory response. The exercise range was chosen to produce a mean level of O2 consumption at or below 50% maximum O2 consumption. Cross-covariant analysis of the fPRBS exercise with breath-by-breath ventilation provided an estimate of the dynamic (impulse) response to exercise, which contained both fast phase 1 and slow phase 2 components. The initial, phase one, hyperpnea occurred within the same breath as the exercise transition and preceded a hypocapnic response. The phase one hyperpnea represented 26% of the total ventilatory response. The secondary, phase 2, hyperpnea was delayed several breaths from the onset of phase 1. It contained slower dynamics and followed a hypercapnic response. Heart rate increased abruptly during phase 1, peaked near the phase 1-to-2 boundary, and then decreased rapidly. The experimental protocol was designed to minimize the subjective response and provide an adequate stimulus for the faster time constants. Results obtained from these experiments were consistent with a nonhumoral induced phase 1 exercise hyperpnea.


The protective effect of thymoquinone against sepsis syndrome morbidity and mortality in mice.
Sepsis and septic shock are life threatening complications and most common cause of death in intensive care units. Thymoquinone, a constituent of Nigella sativa (black seed), holds exceptional promise as an anti-cancer and anti-inflammatory agent. No evidence has been published, however, whether this compound has a protective effect from sepsis-related morbidity, mortality and associated organ dysfunction. To examine this, two sets of mice (n=12 per group), with parallel control groups, were acutely treated with thymoquinone intraperitoneal injections of 1.0 and 2.0mg/kg body weight, and were subsequently challenged with endotoxin Gram-negative bacteria (LPS O111:B4). In another set of experiments, thymoquinone was administered at doses of 0.75 and 1.0mg/kg/day for three consecutive days prior to sepsis induction with live Escherichia coli. Survival of various groups was computed, and renal, hepatic and sepsis markers were quantified. Thymoquinone reduced mortality by 80-90% and improved both renal and hepatic biomarker profiles. The concentrations of IL-1α with 0.75 mg/kg thymoquinone dose was 310.8 ± 70.93 and 428.3 ± 71.32 pg/ml in the 1mg/kg group as opposed to controls (1187.0 ± 278.64 pg/ ml; P<0.05). Likewise, IL-10 levels decreased significantly with 0.75 mg/kg thymoquinone treatment compared to controls (2885.0 ± 553.98 vs. 5505.2 ± 333.96 pg/ml; P<0.01). Mice treated with thymoquinone also exhibited relatively lower levels of TNF-α and IL-2 (P values=0.1817 and 0.0851, respectively). This study gives strength to the potential clinical relevance of thymoquinone in sepsis-related morbidity and mortality reduction and suggests that human studies should be performed.


Long-term follow-up of patients with chronic hepatitis C treated with α-interferon and ribavirin antiviral therapy: clinical and fibrosis impact of treatment response.
The slow progression of chronic hepatitis C (CHC) infection requires long observation periods to detect clinical changes. We compare the incidence of clinical events, hepatocellular carcinoma (HCC), overall mortality, liver-related mortality, and fibrosis progression between patients with a sustained virological response (SVR) and nonresponders (NR) after a 13-year follow-up period. One hundred and eighty-two CHC patients, who received interferon and ribavirin treatment between 1996 and 2000, were included. Clinical events were evaluated during follow-up. At the end of follow-up, transient elastography was used to assess fibrosis progression. Of the 182 patients, 46.7% (n=85) achieved an SVR. Twenty-seven patients developed hepatic decompensation (one SVR) and 15 developed HCC (three SVR). Twenty-nine patients died (eight SVR). Twelve of the 29 deaths were liver related (two SVR). Independent factors associated with hepatic decompensation were NR to treatment [hazard ratio (HR)=23.35; 95% confidence interval (CI): 2.90-189.25; P=0.003], advanced fibrosis at baseline (HR=9.11; 95% CI: 4.13-20.09), and treatment delay after diagnosis (HR=1.02; 95% CI: 1.00-1.03; P=0.012). Only the latter two were associated with HCC development and liver-related mortality. An assessment of liver fibrosis was performed on 125 patients (66 SVR). Fibrosis values were significantly lower in SVR patients, showing less progression to advanced stages of fibrosis [SVR: 6.6 (2.8); 95% CI: 5.8-7.3] than NR [NR: 14.0 (11.1); 95% CI: 11.1-16.9; P<0.001]. In patients with CHC, SVR is durable and reduces clinical events. The risk of HCC development is lower, but not eliminated. Sustained responders showed fibrosis stabilization or improved fibrosis values.


Reverse Genetics of Zika Virus Using a Bacterial Artificial Chromosome.
Zika virus (ZIKV) is a mosquito-borne member of the Flaviviridae family that has become a global threat to human health. Although ZIKV has been known to circulate for decades causing mild febrile illness, the more recent ZIKV outbreaks in the Americas and the Caribbean have been associated with severe neurological disorders and congenital abnormalities. The development of ZIKV reverse genetics approaches have allowed researchers to address key questions on the biology of ZIKV by genetically engineering infectious recombinant (r)ZIKV. This has resulted in a better understanding of the biology of ZIKV infections, including viral pathogenesis, molecular mechanisms of viral replication and transcription, or the interaction of viral and host factors, among others aspects. In addition, reverse genetics systems have facilitated the identification of anti-ZIKV compounds and the development of new prophylactic approaches to combat ZIKV infections. Different reverse genetics strategies have been implemented for the recovery of rZIKV. All these reverse genetics systems have faced and overcome multiple challenges, including the viral genome size, the toxicity of viral sequences in bacteria, etc. In this chapter we describe the generation of a ZIKV full-length complementary (c)DNA infectious clone based on the use of a bacterial artificial chromosome (BAC) and the experimental procedures for the successful recovery of rZIKV. Importantly, the protocol described in this chapter provides a powerful method for the generation of infectious clones of other flaviviruses with genomes that have stability problems during bacterial propagation.


Is it possible to check microcomponent coatings?
Optical microcomponents are increasingly used in laser optical systems because of their many and novel industrial applications. These components are coated in order to enhance their optical performance, but optical characterizations are very difficult due to the shapes and small size. Thus, to perform this kind of measurement, special devices are needed. It is difficult to check component optical responses after manufacturing. Thus a new method, developed by the French Atomic Energy and Alternative Energies Commission, is proposed to fill this gap.


Voice onset time versus articulatory modeling for stop consonants.
Voice onset time (VOT) was developed as a parameter for the pattern-playback speech synthesizer developed approximately 50 years ago, in order to generate the acoustic effects of voiced-unvoiced differences in English stop consonants. However, problems arose when the VOT parameter was used to define stops in actual spoken language, to replace aerodynamic and physiological parameters. A representative physiological model from the same time-period that avoided these problems is sketched. In this model, the manner-of-articulation of a stop is determined by the duration, timing, and extent of laryngeal, articulatory, and respiratory gestures. It is concluded that the term voice onset time should be used only as a parameter in speech synthesis, as originally intended, and not for the analysis of actual speech.


Aspiration-related pulmonary syndromes.
Aspiration of foreign matter into the airways and lungs can cause a wide spectrum of pulmonary disorders with various presentations. The type of syndrome resulting from aspiration depends on the quantity and nature of the aspirated material, the chronicity, and the host responses. Aspiration is most likely to occur in subjects with a decreased level of consciousness, compromised airway defense mechanisms, dysphagia, gastroesophageal reflux, and recurrent vomiting. These aspiration-related syndromes can be categorized into airway disorders, including vocal cord dysfunction, large airway obstruction with a foreign body, bronchiectasis, bronchoconstriction, and diffuse aspiration bronchiolitis, or parenchymal disorders, including aspiration pneumonitis, aspiration pneumonia, and exogenous lipoid pneumonia. In idiopathic pulmonary fibrosis, aspiration has been implicated in disease progression and acute exacerbation. Aspiration may increase the risk of bronchiolitis obliterans syndrome in patients who have undergone a lung transplant. Accumulating evidence suggests that a causative role for aspiration is often unsuspected in patients presenting with aspiration-related pulmonary diseases; thus, many cases go undiagnosed. Herein, we discuss the broadening spectrum of these pulmonary syndromes with a focus on presenting features and diagnostic aspects.


Microarray-based gene expression profiling suggests adaptation of lung epithelial cells subjected to chronic cyclic strain.
Mechanical strain of the lung tissue is a physiological process that affects the behavior of lung cells. Since recent evidence also suggests alterations in the expression of certain genes as a consequence of mechanotransduction, our study aimed at the analysis of the gene expression profile in lung epithelial cells subjected to chronic cyclic strain. Various human lung epithelial cell lines (A549 as principal adherent cell line and four others) were subjected to cyclic strain (16 % surface distension, 12 min(-1)) in a Strain Cell Culture Device for 24 h. In comparison to static controls, expression analyses were performed by gene microarray and qPCR. Microarray analysis revealed many differences in the gene expression but at moderate levels. Altogether 25 genes were moderately down-regulated (0.86-fold ± 0.06) and 26 genes were up-regulated (1.18-fold ± 0.10) in A549 and the others. Strain-regulated genes often code for transcription factors, such as E2F4 and SRF. qPCR analyses confirmed the up-regulation of both transcription factors and further genes, such as PLAU (urokinase-type plasminogen activator) and S100A4 (S100 protein A4). Moreover, we showed the down-regulations of AGR2 (anterior gradient 2) and LCN2 (lipocalin 2). We identified many genes of which the expression was moderately altered in lung epithelial cells subjected to chronic cyclic strain. Although many moderate changes in the gene expression profile might affect cellular behavior, it also suggests an effective adaptation of cells to mechanical forces in long-term conditions.


Isolation and characterization of Desulfitobacterium frappieri sp. nov., an anaerobic bacterium which reductively dechlorinates pentachlorophenol to 3-chlorophenol.
An anaerobic bacterium, strain PCP-1T (T = type strain), which dechlorinates pentachlorophenol (PCP) to 3-chlorophenol, was isolated from a methanogenic consortium. This organism is a spore-forming rod-shaped bacterium that is nonmotile, asaccharolytic, and Gram stain negative but Gram type positive as determined by electron microscopic observations. Inorganic electron acceptors, such as sulfite, thiosulfate, and nitrate (but not sulfate), stimulate growth in the presence of pyruvate and yeast extract. The optimum pH and optimum temperature for growth are 7.5 and 38 degrees C, respectively. The dechlorination pathway is: PCP-->2,3,4,5-tetrachlorophenol -->3,4,5-trichlorophenol-->3,5-dichlorophenol-->3-chlorophenol. This bacterium dechlorinates several different chlorophenols at ortho, meta, and para positions; exceptions to this are 2,3-dichlorophenol, 2,5-dichlorophenol, 3,4-dichlorophenol, and the monochlorophenols. The time course of PCP dechlorination suggests that two enzyme systems are involved in dehalogenation in strain PCP-1T. One system is inducible for ortho dechlorination, and the second system is inducible for meta and para dechlorinations. A 16S rRNA analysis revealed that strain PCP-1T exhibits 95% homology with Desulfitobacterium dehalogenans JW/IU-DC1, an anaerobic bacterium which can dehalogenate chlorophenols only in ortho positions. These results suggest that strain PCP-1T is a member of a new species and belongs to the recently proposed genus Desulfitobacterium. Strain PCP-1T differs from D. dehalogenans JW/IU-DC1 by its broader range of chlorophenol dechlorination. Strain PCP-1 is the type strain of the new species, Desulfitobacterium frappieri.


Particle and microorganism enumeration data: enabling quantitative rigor and judicious interpretation.
Many of the methods routinely used to quantify microscopic discrete particles and microorganisms are based on enumeration, yet these methods are often known to yield highly variable results. This variability arises from sampling error and variations in analytical recovery (i.e., losses during sample processing and errors in counting), and leads to considerable uncertainty in particle concentration or log(10)-reduction estimates. Conventional statistical analysis techniques based on the t-distribution are often inappropriate, however, because the data must be corrected for mean analytical recovery and may not be normally distributed with equal variance. Furthermore, these statistical approaches do not include subjective knowledge about the stochastic processes involved in enumeration. Here we develop two probabilistic models to account for the random errors in enumeration data, with emphasis on sampling error assumptions, nonconstant analytical recovery, and discussion of counting errors. These models are implemented using Bayes' theorem to yield posterior distributions (by numerical integration or Gibbs sampling) that completely quantify the uncertainty in particle concentration or log(10)-reduction given the experimental data and parameters that describe variability in analytical recovery. The presented approach can easily be implemented to correctly and rigorously analyze single or replicate (bio)particle enumeration data.


Ethical issues in the development of tele-ICUs.
The past two decades have seen a dramatic increase in the use of telemedicine while the information technology revolution has contributed significantly to its popularity. In addition, there has been a recent increase in the use of telemedicine in the intensive care unit (ICU), partially driven by a critical shortage of intensivists. However, the ethical questions raised by the implementation of tele-ICUs have not been adequately considered. In this essay, we will discuss the development of tele-ICUs from the perspective of autonomy, beneficence/non-malificence, justice and professionalism.


Modulating Magnetism in Ferroelectric Polymer-Gated Perovskite Manganite Films with Moderate Gate Pulse Chains.
Most previous attempts on achieving electric-field manipulation of ferromagnetism in complex oxides, such as La0.66Sr0.33MnO3 (LSMO), are based on electrostatically induced charge carrier changes through high-k dielectrics or ferroelectrics. Here, the use of a ferroelectric copolymer, polyvinylidene fluoride with trifluoroethylene [P(VDF-TrFE)], as a gate dielectric to successfully modulate the ferromagnetism of the LSMO thin film in a field-effect device geometry is demonstrated. Specifically, through the application of low-voltage pulse chains inadequate to switch the electric dipoles of the copolymer, enhanced tunability of the oxide magnetic response is obtained, compared to that induced by ferroelectric polarization. Such observations have been attributed to electric field-induced oxygen vacancy accumulation/depletion in the LSMO layer upon the application of pulse chains, which is supported by surface-sensitive-characterization techniques, including X-ray photoelectron spectroscopy and X-ray magnetic circular dichroism. These techniques not only unveil the electrochemical nature of the mechanism but also establish a direct correlation between the oxygen vacancies created and subsequent changes to the valence states of Mn ions in LSMO. These demonstrations based on the pulsing strategy can be a viable route equally applicable to other functional oxides for the construction of electric field-controlled magnetic devices.


Elucidation of fluorine's impact on pKa and in vitro Pgp-mediated efflux for a series of PDE9 inhibitors.
P-Glycoprotein (Pgp)-mediated cellular efflux is recognized as a common challenge in CNS drug discovery. In this study, the influence of replacing a hydrogen atom with fluorine on the pKa and Pgp-mediated efflux is elucidated for a series of PDE9 inhibitors. The PDE9 inhibitors with and without fluorine were synthesized using a novel condensation-oxidation approach, providing access to several analogues, all from the same stereoenriched aldehyde building block. The incorporation of fluorine was found to influence two acid-base functionalities concomitantly, both of which were involved in Pgp-recognition. By methylating the acidic functionality, it was possible to isolate the effect responsible for lowering the Pgp-mediated efflux.


Frequency invariant classification of ultrasonic weld inspection signals.
Automated signal classification systems are finding increasing use in many applications for the analysis and interpretation of large volumes of signals. Such systems show consistency of response and help reduce the effect of variabilities associated with human interpretation. This paper deals with the analysis of ultrasonic NDE signals obtained during weld inspection of piping in boiling water reactors. The overall approach consists of three major steps, namely, frequency invariance, multiresolution analysis, and neural network classification. The data are first preprocessed whereby signals obtained using different transducer center frequencies are transformed to an equivalent reference frequency signal. Discriminatory features are then extracted using a multiresolution analysis technique, namely, the discrete wavelet transform (DWT). The compact feature vector obtained using wavelet analysis is classified using a multilayer perceptron neural network. Two different databases containing weld inspection signals have been used to test the performance of the neural network. Initial results obtained using this approach demonstrate the effectiveness of the frequency invariance processing technique and the DWT analysis method employed for feature extraction.


Risk factors for Salmonella Dublin on dairy farms in Ontario, Canada.
Salmonella Dublin (S. Dublin) is an emerging pathogen on dairy farms in Canada. In Ontario, S. Dublin has been increasingly isolated from diagnostic laboratory samples. The objective of this observational cross-sectional study was to identify management practices associated with herd positivity for S. Dublin. A convenience sample of 100 dairy farms was visited in Ontario, Canada from April to August 2022. Farms were visited once to collect blood samples from 20 heifers between 4 and 24 mo old, sample bulk tank milk, and administer an in-person questionnaire on management practices. An additional bulk tank milk sample was collected before the visit by milk transporters. All bulk tank and serum samples underwent ELISA testing to determine Salmonella Dublin positivity (≥35% percent positivity on ELISA). Of the 1,990 heifers sampled, 44 (2.2%) animals were seropositive for S. Dublin. At least one seropositive heifer was identified on 24% of participating farms. Based on the bulk tank milk samples collected during both sampling periods, 4% of farms were positive for S. Dublin. Overall, of the 100 farms visited, 25% were classified as S. Dublin positive, meaning at least one serum or bulk tank sample was interpreted as positive. A multivariable logistic regression model identified 5 factors associated with herd-level positivity for S. Dublin. Specifically, introducing purchased animals within the last 2 years increased the likelihood that farms were positive for S. Dublin (odds ratio (OR) = 4.6). Farms that had at least one animal leave the premises for a cattle show, embryo collection center, or loan to another farm and return within the last 2 years were also at a higher risk for S. Dublin (OR = 4.9). Farms that removed manure from the surface of bedding in calving pens twice per month or after every calving were at greater risk for S. Dublin than farms that removed manure less frequently (OR = 8.5). Farms that added bedding material to calving areas once or twice weekly were at lower risk for S. Dublin compared with farms that added bedding less than once weekly (OR = 0.1). In addition, farms that kept 3 cows or less per pen in the calving area were at lower risk for S. Dublin. Test positivity for S. Dublin among Ontario dairy farms sampled is high, and dairy producers should consider avoiding management practices that are associated with an increased risk of S. Dublin infection.


Analysis of FOXG1 Is Highly Recommended in Male and Female Patients with Rett Syndrome.
We screened a cohort of 5 male and 20 female patients with a Rett spectrum disorder for mutations in the coding region of FOXG1, previously shown to cause the congenital variant of Rett syndrome. Two de novo mutations were identified. The first was a novel missense mutation, p.Ala193Thr (c.577G>A), in a male patient with congenital Rett syndrome, and the second was the p.Glu154GlyfsX301 (c.460dupG) truncating mutation in a female with classical Rett syndrome, a mutation that was previously reported in an independent patient. The overall rate of FOXG1 mutations in our cohort is 8%. Our findings stress the importance of FOXG1 analysis in male patients with Rett syndrome and in female patients when mutations in the MECP2 and CDKL5 genes have been excluded.


Apolipoprotein E polymorphism determined by restriction enzyme analysis of DNA amplified by polymerase chain reaction: convenient alternative to phenotyping by isoelectric focusing.
Three common alleles determine six apolipoprotein E (apo E) phenotypes that are associated with variations in serum cholesterol in the population. This genetic variation results from single nucleotide alterations at two DNA loci encoding the amino acid residues 112 and 158 of apo E. We compared results of apo E phenotyping carried out by isoelectric focusing with those of apo E genotyping accomplished by direct DNA analysis. In the latter, the target DNA was amplified by the polymerase chain reaction (PCR) and subsequently analyzed by digestion with the restriction enzyme Hha I, followed by polyacrylamide gel electrophoresis of the cleavage products. With one exception, these two techniques yielded similar results from all 40 samples tested. In addition, a rare variant form of apo E (phenotype E1) was analyzed separately and incorrectly diagnosed as E2 by the Hha I digestion method; the anticipated mutation in the codon 127 was, however, confirmed by demonstration of a new Taq I restriction site in this variant gene. These data confirm that the common isoforms of apo E usually arise from genetic variation of the codons 112 and 158 and demonstrate the feasibility of the PCR technique in apo E genotyping.


New advances in hematopoietic cell transplantation.
The present review highlights sentinel work published since 2006 on the definition of the transplantation barrier and the elucidation of cytokine and immune response gene variation in defining posttransplant risks. Recent work has defined the relative importance of matching for the classical human leukocyte antigen (HLA) HLA-A, HLA-B, HLA-C, DRB1, DQB1 genes, and the importance of additive effects of multilocus disparity. This work provides a new framework for donor identification and extends the use of single locus HLA-DQB1 mismatched donors without compromising the success of the transplant. New data demonstrate that permissible class I mismatches may be defined by donor-recipient mismatching at certain residues. The concept that the extended HLA haplotype carries undetected but functional variation provides an approach for mapping novel transplantation determinants, and a means to further improve the clinical results of transplantation from HLA-matched unrelated donors. Finally, the role of sequence variation in immune response and cytokine genes provides a means for assessing risks for a given transplant recipient and may aid in the planning of the transplant procedure. Optimizing the results of unrelated donor transplantation requires an understanding of risks associated with variation of HLA genes within the major histocompatibility complex, and of genes that participate in the immune response and inflammatory pathways.


Assessment of changes following en-masse retraction with mini-implants anchorage compared to two-step retraction with conventional anchorage in patients with class II division 1 malocclusion: a randomized controlled trial.
No randomized controlled trial has tried to compare treatment outcomes between the sliding en-masse retraction of upper anterior teeth supported by mini-implants and the two-step sliding retraction technique employing conventional anchorage devices. To evaluate skeletal, dental, and soft tissue changes following anterior teeth retraction. Parallel-groups randomized controlled trial on patients with class II division 1 malocclusion treated at the University of Al-Baath Dental School in Hamah, Syria between July 2011 and May 2013. One hundred and thirty-three patients with an upper dentoalveolar protrusion were evaluated and 80 patients fulfilled the inclusion criteria. Randomization was performed using computer-generated tables; allocation was concealed using sequentially numbered opaque and sealed envelopes. Fifty-six participants were analysed (mean age 22.34 ± 4.56 years). They were randomly distributed into two groups with 28 patients in each group (1:1 allocation ratio). Following first premolar extraction, space closure was accomplished using either the en-masse technique with mini-implants or the two-step technique with transpalatal arches (TPAs). The antero-posterior displacements of upper incisal edges and upper first molars were measured on lateral cephalograms at three assessment times. Assessor blinding was employed. A bodily retraction (-4.42 mm; P < 0.001) with a slight intrusion (-1.53 mm; P < 0.001) of the upper anterior teeth was achieved in the mini-implants group, whereas upper anterior teeth retraction was achieved by controlled palatal tipping in the TPA group. When retracting anterior teeth in patients with moderate to severe protrusion, the en-masse retraction based on mini-implants anchorage gave superior results compared to the two-step retraction based on conventional anchorage in terms of speed, dental changes, anchorage loss, and aesthetic outcomes.


5HT(2A) and 5HT(2B) receptors contribute to serotonin-induced vascular dysfunction in diabetes.
Although 5HT(2A) receptors mediate contractions of normal arteries to serotonin (5HT), in some cardiovascular diseases, other receptor subtypes contribute to the marked increase in serotonin contractions. We hypothesized that enhanced contractions of arteries from diabetics to 5HT are mediated by an increased contribution from multiple 5HT receptor subtypes. We compared responses to selective 5HT receptor agonists and expression of 5HT receptor isoforms (5HT(1B), 5HT(2A), and 5HT(2B)) in aorta from nondiabetic (ND) compared to type 2 diabetic mice (DB, BKS.Cg-Dock7(m)+/+Lepr(db)/J). 5HT, 5HT(2A) (TCB2 and BRL54443), and 5HT(2B) (norfenfluramine and BW723C86) receptor agonists produced concentration-dependent contractions of ND arteries that were markedly increased in DB arteries. Neither ND nor DB arteries contracted to a 5HT(1B) receptor agonist. MDL11939, a 5HT(2A) receptor antagonist, and LY272015, a 5HT(2B) receptor antagonist, reduced contractions of arteries from DB to 5HT more than ND. Expression of 5HT(1B), 5HT(2A), and 5HT(2B) receptor subtypes was similar in ND and DB. Inhibition of rho kinase decreased contractions to 5HT and 5HT(2A) and 5HT(2B) receptor agonists in ND and DB. We conclude that in contrast to other cardiovascular diseases, enhanced contraction of arteries from diabetics to 5HT is not due to a change in expression of multiple 5HT receptor subtypes.


Regulation of enzyme turnover during tissue differention. Studies on the effects of hormones on the turnover of fatty acid synthetase in rabbit mammary gland in organ culture.
1. Explants of mammary gland from mid-pregnant rabbits were cultured with insulin, prolactin and cortisol. 2. Antibodies raised to fatty acid synthetase were used to measure the amount as well as the rate of synthesis and the rate of degradation of the enzyme in the explants over defined periods in organ culture. These measurements were also made after the hormones had been removed from the culture medium. The changes which occur in the activity of fatty acid synthetase are due to changes in the amount of the enzyme present. They are not due to activation or inactivation of the enzyme. 3. The rate of lipogenesis (measured from [1-14C]acetate) in the explants during culture varies independently of the amount of fatty acid synthetase both in the presence and after removal of the hormones. Hence the amount of fatty acid synthetase does not limit lipogenesis. The proportion of medium-chain fatty acids C8:0 and C10:0 (which are characteristic of rabbit milk) synthesized by the explants in the presence of hormones increases at about the same rate as the amount of fatty acid synthetase present. However, when hormones are removed from the medium the proportion of these acids synthesized declines as rapidly as the rate of lipogenesis and not as the amount of fatty acid synthetase presen. 4. The rates of synthesis of fatty acid synthetase and of the total particulate-free supernatant protein in the explants were compared by measuring the incorporation of L-[U-14C]leucine into the protein of the explants. These rates increase by 5-fold and 3.6-fold respectively when explants are cultured with hormones, and they then reach approximately constant rates. When the hormones are removed there is a rapid fall in the rate of synthesis of fatty acid synthetase and of the total particulate-free supernatant protein to values which are similar to those obtained with freshly prepared explanted tissue. 5. In unstimulated explants fatty acid synthetase appears to be degraded with a half-life of 15-21h. During the hormonally stimulated differentiation of the tissue the rate of degradation of the enzyme is considerably decreased or is switched off completely. After the amount of fatty acid synthetase has increased to a maximum the enzyme complex is again degraded with a half-life of 23-29h. The removal of hormones after the explants have been hormonally stimulated for different times results in an increase in the rate of degradation of fatty acid synthetase. However, this increase only occurs if degradation was previously proceeding at a considerably decreased rate. The degradation of the total particulate-free supernatant protein continues throughout the period of differentiation of the explant tissue in culture. It appears to be somewhat decreased during the period of rapid maturation of the tissue during culture.


Laparoscopic surgery for rectal cancer: short-term benefits and oncologic outcomes using more than one technique.
Several minimally invasive techniques have now been described for rectal cancer resection. Current outcome data for these approaches from high volume, single institutions remain limited. Our aim was to review outcomes in patients undergoing minimally invasive surgery for rectal cancer at our institution in the current era. A retrospective analysis was done to assess short-term benefits and oncologic outcomes in patients undergoing minimally invasive surgery for rectal cancer between 2004 and 2007. One-hundred consecutive patients (61 men, median age 62) with a median follow-up of 1.8 years were identified. Sixty-seven had hand-assisted laparoscopic surgery (HALS), while 33 were done laparoscopic-assisted (LA). Seventy-two patients underwent anterior resection, 27 an abdominal perineal resection, and 1 a total proctocolectomy. Tumor stage was stage 1 (21%), stage 2 (17%), stage 3 (56%), and stage 4 (6%). A median of 16 lymph nodes, a median 3.4 cm distal margin, and a 99% negative circumferential margin was achieved. The 3-year disease-free and overall survivals were 86.2 and 94.5%, respectively. Three cases required conversion. Median time to both diet and first bowel movement was 3 days, and median length of stay was 5 days. Length of stay, time to soft diet, incision length, and pain scores were less using a LA approach compared to HALS (P < 0.01). Overall morbidity was 26% with no mortality. Both minimally invasive techniques used achieved excellent oncologic results in patients with rectal cancer. The LA approach had slightly better short-term outcomes.


A Phase II pilot trial to evaluate safety and efficacy of ferroquine against early Plasmodium falciparum in an induced blood-stage malaria infection study.
Ferroquine (SSR97193) is a candidate anti-malarial currently undergoing clinical trials for malaria. To better understand its pharmacokinetic (PK) and pharmacodynamic (PD) parameters the compound was tested in the experimentally induced blood stage malaria infection model in volunteers. Male and non-pregnant female aged 18-50 years were screened for this phase II, controlled, single-centre clinical trial. Subjects were inoculated with ~1800 viable Plasmodium falciparum 3D7A-infected human erythrocytes, and treated with a single-dose of 800 mg ferroquine. Blood samples were taken at defined time-points to measure PK and PD parameters. The blood concentration of ferroquine and its active metabolite, SSR97213, were measured on dry blood spot samples by ultra-performance liquid chromatography with tandem mass spectrometry (LC-MS/MS). Parasitaemia and emergence of gametocytes were monitored by quantitative PCR. Safety was determined by recording adverse events and monitoring clinical laboratory assessments during the course of the study. Eight subjects were enrolled into the study, inoculated with infected erythrocytes and treated with 800 mg ferroquine. Ferroquine was rapidly absorbed with maximal exposure after 4-8 and 4-12 h exposure for SSR97213. Non-compartmental PK analysis resulted in estimates for half-lives of 10.9 and 23.8 days for ferroquine and SSR97213, respectively. Parasite clearance as reported by parasite reduction ratio was 162.9 (95 % CI 141-188) corresponding to a parasite clearance half-life of 6.5 h (95 % CI: 6.4-6.7 h). PK/PD modelling resulted in a predicted minimal parasiticidal concentration of 20 ng/mL, and the single dosing tested in this study was predicted to maintain an exposure above this threshold for 454 h (37.8 days). Although ferroquine was overall well tolerated, transient elevated transaminase levels were observed in three subjects. Paracetamol was the only concomitant treatment among the two out of these three subjects that may have played a role in the elevated transaminases levels. No clinically significant ECG abnormalities were observed. The parameters and PK/PD model derived from this study pave the way to the further rational development of ferroquine as an anti-malarial partner drug. The safety of ferroquine has to be further explored in controlled human trials. Trial registration anzctr.org.au (registration number: ACTRN12613001040752), registered 18/09/2013.