The impact of intracranial carotid artery calcification on the development of thrombolysis-induced intracerebral hemorrhage.
We aimed to assess whether intracranial carotid artery calcification (ICAC) evident on head computed tomography is a risk factor for symptomatic intracerebral hemorrhage (sICH) following tissue plasminogen activator (tPA) treatment for acute stroke. We classified 297 consecutive patients into 2 groups (no to mild ICAC and moderate to severe ICAC) according to ICAC severity. Outcome measures included detection of intracerebral hemorrhage and assessment using a modified Rankin scale (mRS) at 1 month and 1 year after stroke. ICH (any type) was significantly more common in patients with moderate to severe ICAC than in patients with no to mild ICAC (22.5% versus 12%; relative risk [RR], 1.67; 95% confidence interval [CI], 1.1-2.5; P<.05). The moderate to severe ICAC group tended to have a higher percentage of sICH, but this association was not statistically significant (RR, 1.57; 95% CI, .75-3.3, P>.05). Multivariate adjusted regression analysis revealed that moderate to severe ICAC was an independent risk factor for ICH following tPA treatment (odds ratio, 2.52; 95% CI, 1.07-5.94; P=.04). Dependent functional outcome (mRS score 3-6) at 1-month and 1-year follow-up was significantly associated with moderate to severe ICAC (RR, 1.56; 95% CI, 1.06-2.27; and RR, 1.56; 95% CI, 1.06-2.33; P<.05). However, ICAC was not an independent factor of functional dependency at 1-month and 1-year follow-up in the final multivariate regression model. A significantly higher percentage of patients with moderate to severe ICAC developed ICH following tPA administration for stroke. ICAC severity is an independent risk factor for ICH events. ICAC severity can help predict short-term and long-term functional dependency in tPA-treated patients, although this can be confounded by other cardiovascular risk factors and stroke severity.


Involvement of sperm sulphatases in early sperm-zona interactions in the hamster.
Sulphatase preparations from Abalone entrails, the limpet Patella vulgata and ox liver, as well as artificial substrates for these enzymes, were used in the hamster in vitro fertilization system to study the possible roles of sperm sulphatases in sperm-zona pellucida interactions. p-nitrophenyl sulphate, p-nitrocatechol sulphate, ascorbate 2-sulphate, as well as D-glucopyranose and D-galactopyranose, both sulphated at the 3, 4 or 6 position but not the 2 position, inhibited fertilization in a dose-dependent manner. Sperm-egg fusion was not inhibited by the substrates used and eggs pre-treated with sulphates could readily be fertilized. Sperm motility and therefore viability was unaffected by inhibitory concentrations of substrates as determined by rhodamine 123 labelling of motile spermatozoa. Acrosomal integrity of rhodamine-labelled spermatozoa was assessed and found to be unaffected by inhibitory levels of substrates. Fertilization was inhibited by high concentrations of the two molluscan sulphatases but not by purified ox liver sulphatase. Pre-treatment of eggs with these enzymes did not prevent fertilization. Long-term exposure of hamster oocytes to N-acetylhexosaminidase or limpet sulphatase caused thinning and distension of the zona pellucida but these changes were not observed with the ox liver sulphatase. The results suggest that a glycosulphatase is probably released from hamster spermatozoa during sperm-egg adhesion and, or, penetration. If sperm-egg adhesion molecules are sulphated, the commercially available sulphatases would be unsuitable for their characterization.


Optimal number of features as a function of sample size for various classification rules.
Given the joint feature-label distribution, increasing the number of features always results in decreased classification error; however, this is not the case when a classifier is designed via a classification rule from sample data. Typically (but not always), for fixed sample size, the error of a designed classifier decreases and then increases as the number of features grows. The potential downside of using too many features is most critical for small samples, which are commonplace for gene-expression-based classifiers for phenotype discrimination. For fixed sample size and feature-label distribution, the issue is to find an optimal number of features. Since only in rare cases is there a known distribution of the error as a function of the number of features and sample size, this study employs simulation for various feature-label distributions and classification rules, and across a wide range of sample and feature-set sizes. To achieve the desired end, finding the optimal number of features as a function of sample size, it employs massively parallel computation. Seven classifiers are treated: 3-nearest-neighbor, Gaussian kernel, linear support vector machine, polynomial support vector machine, perceptron, regular histogram and linear discriminant analysis. Three Gaussian-based models are considered: linear, nonlinear and bimodal. In addition, real patient data from a large breast-cancer study is considered. To mitigate the combinatorial search for finding optimal feature sets, and to model the situation in which subsets of genes are co-regulated and correlation is internal to these subsets, we assume that the covariance matrix of the features is blocked, with each block corresponding to a group of correlated features. Altogether there are a large number of error surfaces for the many cases. These are provided in full on a companion website, which is meant to serve as resource for those working with small-sample classification. For the companion website, please visit http://public.tgen.org/tamu/ofs/ firstname.lastname@example.com.


Measuring Geometric Phase without Interferometry.
A simple noninterferometric approach for probing the geometric phase of a structured Gaussian beam is proposed. Both the Gouy and Pancharatnam-Berry phases can be determined from the intensity distribution following a mode transformation if a part of the beam is covered at the initial plane. Moreover, the trajectories described by the centroid of the resulting intensity distributions following these transformations resemble those of ray optics, revealing an optical analogue of Ehrenfest's theorem associated with changes in the geometric phase.


[Effect of flunarizine on healing and necrosis of experimental autografts in the rat].
The influence of flunarizine on flap necrosis has been studied in rats by means of an abdominal neurovascular island skin flap model. Four groups of six rats were treated in a pretreatment phase of one week and a medication phase of four weeks with either flunarizine (2.5 mg/kg) or a placebo. Rats with flunarizine treatment over the whole period showed a significantly faster healing and less necrosis than the placebo group. The group without flunarizine pretreatment showed similar results as the control group at the first week after surgery, but from the second week on, the flunarizine effect became manifest. The skin graft model clearly demonstrates the beneficial effects of flunarizine in the prevention of tissue necrosis.


Genetics of hypertension. Current status.
Genetics of hypertension is complex with no known single gene playing a major role, but rather many genes each with mild effects reacting to different environmental stimuli contribute to blood pressure. The heritable component of blood pressure has been documented in familial and twin studies suggesting that 30%-50% of the variance of blood pressure readings are attributable to genetic heritability and about 50% to environmental factors. Early studies in hypertension identified specific enzymes, channels and receptors implicating sodium handling in the regulation of blood pressure including genes involved with the renin-angiotensin-aldosterone system controlling blood pressure and salt-water homeostasis, proteins in hormonal regulation of blood pressure (enzymes and receptors of the mineralo- and glucocorticoid pathways) and proteins coded by genes involved in the structure and/or regulation of vascular tone (endothelins and their receptors). The field of molecular genetics has revolutionized the study of hypertension by identifying single gene syndromes or Mendelian forms and several candidate genes for blood pressure variance. Genes have been localized to at least 20 chromosome regions. For example, recent genome-wide association studies (GWAS) of common genetic variants found 13 single nucleotide polymorphisms (SNPs) or variants in systolic and 20 for diastolic blood pressure readings representing different genes and genetic heterogeneity. Further understanding of the genetics of hypertension will require the use of advances in bioinformatics tools and genetic technology [e.g., SNP, exon and noncoding (micro) RNA arrays]. New approaches will allow for identification of not only single genes, but other interacting genes contributing to hypertension by merging multiple genetic data sets (structural and functional) from individuals with hypertension and development of new molecular targets for study and treatment.


Incidence of dementia in a representative British sample.
In a four-year follow-up study of 1042 elderly people (aged 65 years or older), randomly sampled from the community, levels of dementia were assessed using a two-phase case-finding procedure (screening followed by clinical interview) among survivors. Clinical information on those not reinterviewed was provided by death certificates, hospital case notes, or postal questionnaires. The weighted four-year cumulative incidence of dementia was 3.7% (95% confidence intervals: 2.4%-5.0%), with age-specific rates of 0.9%, 2.8%, 5.2%, 9.0%, and 8.7% for the age groups 65-69, 70-74, 75-79, 80-84, and 85-89 years respectively. While consistent with data from other British regions, it remains likely that these rates underestimate true incidence.


Proximal tibial varus osteotomy. Indications, technique, and five to twenty-one-year results.
Although high tibial osteotomy has been proved effective for the treatment of painful osteoarthritis of the medial compartment of the knee, the role of proximal tibial varus osteotomy for the treatment of painful osteoarthritis of the lateral compartment still remains controversial. From 1974 to 1993, we performed proximal tibial varus osteotomy for the treatment of osteoarthritis of the lateral compartment of the knee in thirty-six consecutive patients. The procedure consisted of a proximal lateral opening-wedge varus osteotomy of the tibia with use of corticocancellous bone grafts from the iliac crest. The valgus deformity was posttraumatic in twenty-three patients, followed a lateral meniscectomy in five, was due to overcorrection of a varus deformity in four, and was idiopathic in four. The preoperative valgus deformity averaged 11.6 degrees (range, 4 degrees to 22 degrees ). At a mean of eleven years (range, five to twenty-one years) after the operation, the clinical results for thirty-four of the thirty-six patients were analyzed. None of the patients had severe progression of the osteoarthritis after the osteotomy, and none had a meaningful loss in the range of motion of the knee joint. A superficial wound infection developed in one patient, and another patient had thrombophlebitis. Three patients (9%) had a transient palsy of the peroneal nerve. According to the system of Insall et al., the mean knee score was 84 points (range, 54 to 99 points). According to the knee score described by Lysholm and Gillquist, the subjective result was excellent in nine patients (26%), good in twenty-one (62%), fair in three (9%), and poor in one (3%). We concluded that when the indications outlined in this study are followed and our opening-wedge technique is used, a proximal lateral opening-wedge varus osteotomy of the tibia is a good alternative for the treatment of isolated osteoarthritis of the lateral compartment of the knee. High accuracy in preoperative planning, based on a slight overcorrection, is important to prevent failure.


Linking sight and sound: fMRI evidence of primary auditory cortex activation during visual word recognition.
We describe two studies that used repetition priming paradigms to investigate brain activity during the reading of single words. Functional magnetic resonance images were collected during a visual lexical decision task in which nonword stimuli were manipulated with regard to phonological properties and compared to genuine English words. We observed a region in left-hemisphere primary auditory cortex linked to a repetition priming effect. The priming effect activity was observed only for stimuli that sound like known words; moreover, this region was sensitive to strategic task differences. Thus, a brain region involved in the most basic aspects of auditory processing appears to be engaged in reading even when there is no environmental oral or auditory component.


Mu-opioid receptor trafficking on inhibitory synapses in the rat brainstem.
Whole-cell recordings were made from identified gastric-projecting rat dorsal motor nucleus of the vagus (DMV) neurons. The amplitude of evoked IPSCs (eIPSCs) was unaffected by perfusion with met-enkephalin (ME) or by mu-, delta-, or kappa-opioid receptor selective agonists, namely D-Ala2-N-Me-Phe4-Glycol5-enkephalin (DAMGO), cyclic [D-Pen2-D-Pen5]-enkephalin, or trans-3,4-dichloro-N-methyl-N-[2-(1-pyrolytinil)-cyclohexyl]-benzeneacetamide methane sulfonate (U50,488), respectively. Brief incubation with the adenylate cyclase activator forskolin or the nonhydrolysable cAMP analog 8-bromo-cAMP, thyrotropin releasing hormone, or cholecystokinin revealed the ability of ME and DAMGO to inhibit IPSC amplitude; this inhibition was prevented by pretreatment with the mu-opioid receptor (MOR1) selective antagonist D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2. Conversely, incubation with the adenylate cyclase inhibitor dideoxyadenosine, with the protein kinase A (PKA) inhibitor N-[2-(p-Bromocinnamyl-amino)ethyl]-5-isoquinolinesulfonamide dihydrochloride (H89), or with the Golgi-disturbing agent brefeldin A, blocked the ability of forskolin to facilitate the inhibitory actions of ME. Immunocytochemical experiments revealed that under control conditions, MOR1 immunoreactivity (MOR1-IR) was colocalized with glutamic acid decarboxylase (GAD)-IR in profiles apposing DMV neurons only after stimulation of the cAMP-PKA pathway. Pretreatment with H89 or brefeldin A or incubation at 4 degrees C prevented the forskolin-mediated insertion of MOR1 on GAD-IR-positive profiles. These results suggest that the cAMP-PKA pathway regulates trafficking of mu-opioid receptors into the cell surface of GABAergic nerve terminals. By consequence, the inhibitory actions of opioid peptides in the dorsal vagal complex may depend on the state of activation of brainstem vagal circuits.


Protective effect of bioflavonoid myricetin enhances carbohydrate metabolic enzymes and insulin signaling molecules in streptozotocin-cadmium induced diabetic nephrotoxic rats.
Diabetic nephropathy is the kidney disease that occurs as a result of diabetes. The present study was aimed to evaluate the therapeutic potential of myricetin by assaying the activities of key enzymes of carbohydrate metabolism, insulin signaling molecules and renal function markers in streptozotocin (STZ)-cadmium (Cd) induced diabetic nephrotoxic rats. After myricetin treatment schedule, blood and tissue samples were collected to determine plasma glucose, insulin, hemoglobin, glycosylated hemoglobin and renal function markers, carbohydrate metabolic enzymes in the liver and insulin signaling molecules in the pancreas and skeletal muscle. A significant increase of plasma glucose, glycosylated hemoglobin, urea, uric acid, creatinine, blood urea nitrogen (BUN), urinary albumin, glycogen phosphorylase, glucose-6-phosphatase, and fructose-1,6-bisphosphatase and a significant decrease of plasma insulin, hemoglobin, hexokinase, glucose-6-phosphate dehydrogenase, glycogen and glycogen synthase with insulin signaling molecule expression were found in the STZ-Cd induced diabetic nephrotoxic rats. The administration of myricetin significantly normalizes the carbohydrate metabolic products like glucose, glycated hemoglobin, glycogen phosphorylase and gluconeogenic enzymes and renal function markers with increase insulin, glycogen, glycogen synthase and insulin signaling molecule expression like glucose transporter-2 (GLUT-2), glucose transporter-4 (GLUT-4), insulin receptor-1 (IRS-1), insulin receptor-2 (IRS-2) and protein kinase B (PKB). Based on the data, the protective effect of myricetin was confirmed by its histological annotation of the pancreas, liver and kidney tissues. These findings suggest that myricetin improved carbohydrate metabolism which subsequently enhances glucose utilization and renal function in STZ-Cd induced diabetic nephrotoxic rats.


In mice with type 2 diabetes, a vascular endothelial growth factor (VEGF)-activating transcription factor modulates VEGF signaling and induces therapeutic angiogenesis after hindlimb ischemia.
Peripheral arterial disease is a major complication of diabetes. The ability to promote therapeutic angiogenesis may be limited in diabetes. Type 2 diabetes was induced by high-fat feeding C57BL/6 mice (n = 60). Normal chow-fed mice (n = 20) had no diabetes. Mice underwent unilateral femoral artery ligation and excision. A plasmid DNA encoded an engineered transcription factor designed to increase vascular endothelial growth factor expression (ZFP-VEGF). On day 10 after the operation, the ischemic limbs received 125 microg ZFP-VEGF plasmid or control. Mice were killed 3, 10, or 20 days after injection (n = 10/group, at each time point). Limb blood flow was measured by laser Doppler perfusion imaging. VEGF mRNA expression was examined by real-time PCR. VEGF, Akt, and phospho-Akt protein were measured by enzyme-linked immunosorbent assay. Capillary density, proliferation, and apoptosis were assessed histologically. Compared with normal mice, mice with diabetes had greater VEGF protein, reduced phospho-Akt-to-Akt ratio before ligation, and an impaired perfusion recovery after ligation. At 3 and 10 days after injection, in mice with diabetes, gene transfer increased VEGF expression and signaling. At later time points, gene transfer resulted in better perfusion recovery. Gene transfer with ZFP-VEGF was able to promote therapeutic angiogenesis mice with type 2 diabetes.


Partnering for quality.
This article discusses efforts being made by multiple stakeholders to improve quality and safety in intensive care units (ICUs). The Leapfrog Group (Washington, DC), a consortium of purchasers, is reaching for quality by implementing safety standards. One standard, staffing ICUs with intensivists, is associated with a 30% reduction in hospital mortality and a 40% reduction in ICU mortality. While Leapfrog is addressing quality improvements on a structural level, JCAHO is developing empiric measures for ICU quality, which are being beta tested, to complement Leapfrogs' efforts. JCAHO used rigorously defined processes to develop and refine these measures. These measures include deep venous thrombosis prophylaxis, peptic ulcer disease prophylaxis, catheter-related blood stream infections, mortality and length of stay-APACHE III risk adjustment. Finally, ICU providers at Hopkins implemented an 8-step unit-based safety program. This comprehensive program is run at the unit level, works to improve safety culture, addresses staff safety concerns, partners hospital leaders with unit workers, helps staff implement improvement initiatives, uses rigorous data collection methods, and easily disseminated in other clinical areas. Providers can frame their efforts after 3 fundamental concepts from these initiatives: create a culture that supports quality and safety, standardize work processes by reducing complexity and using independent redundancy, and automate. The take home message is, when we come to work every day, we must be committed to quality and safety, and lead our teams to do the same.


A point prevalence survey of health care-associated infections in Canadian pediatric inpatients.
Health care-associated infections (HAIs) cause considerable morbidity and mortality to hospitalized patients. The objective of this point prevalence study was to assess the burden of HAIs in the Canadian pediatric population, updating results reported from a similar study conducted in 2002. A point prevalence survey of pediatric inpatients was conducted in February 2009 in 30 pediatric or combined adult/pediatric hospitals. Data pertaining to one 24-hour period were collected, including information on HAIs, microorganisms isolated, antimicrobials prescribed, and use of additional (transmission based) precautions. The following prevalent infections were included: pneumonia, urinary tract infection, bloodstream infection, surgical site infection, viral respiratory infection, Clostridium difficile infection, viral gastroenteritis, and necrotizing enterocolitis. One hundred eighteen patients had 1 or more HAI, corresponding to a prevalence of 8.7% (n = 118 of 1353, 95% confidence interval: 7.2-10.2). Six patients had 2 infections. Bloodstream infections were the most frequent infection in neonates (3.0%), infants (3.1%), and children (3.5%). Among all patients surveyed, 16.3% were on additional precautions, and 40.1% were on antimicrobial agents, whereas 40.7% of patients with a HAI were on additional precautions, and 89.0% were on antimicrobial agents. Overall prevalence of HAI in 2009 has remained similar to the prevalence reported from 2002. The unchanged prevalence of these infections nonetheless warrants continued vigilance on their prevention and control.


Strengthening the HIV Care Continuum in the Dominican Republic: Application of a Triadic Implementation Framework to Meet the UNAIDS 90-90-90 Treatment Goal.
Innovative empirical frameworks to evaluate progress in efforts addressing HIV treatment and prevention barriers in resource-limited areas are sorely needed to achieve the UNAIDS 90-90-90 goal (90% diagnosed, 90% on treatment, and 90% virally suppressed). A triadic implementation framework (TIF) is a comprehensive conceptual tool for (1) monitoring attrition, (2) evaluating operational programs, and (3) measuring the impact of specific implementation goals within the care continuum. TIF will assess the effects of enhanced programs on adherence and virologic suppression within the HIV care continuum at a regional clinic in the Dominican Republic (Clínica de Familia La Romana [CFLR]) and its program serving high-risk, migratory batey (sugarcane cultivation) communities. A retrospective cohort study completed during 2015 collected deidentified data from a CFLR chart review of adult HIV patients diagnosed in 2013. The results were quantitatively analyzed and compared to 2011 cohort data. In 2013, 310 patients were diagnosed HIV positive. The results demonstrated 73% enrolling in care, 28% adhering to care, and 16% achieving viral load suppression. Engagement increased across all steps of the care continuum compared to a 2011 cohort, culminating in a significant increase in undetectable viral load from 4% to 16% (p < 0.001). The batey program showed significant increases in patient enrollment compared to the 2011 cohort (p < 0.001). Meeting the UNAIDS 90-90-90 goal requires enhanced services in high-burden, resource-limited regions. CFLR employs TIF to assess progress and programmatic areas in need of strengthening. Data suggest enhanced CFLR services improve outcomes. Given improvements, maintenance and expansion of similar programs are warranted to achieve the 90-90-90 goal.


Different mechanisms of decreased drug accumulation in doxorubicin and mitoxantrone resistant variants of the MCF7 human breast cancer cell line.
We selected two drug resistant variants of the MCF7 human breast cancer cell line by chronic in vitro exposure to doxorubicin (MCF7/D40 cell line) and mitoxantrone (MCF7/Mitox cell line), respectively. The cell lines are similar in growth characteristics including doubling time, DNA synthetic phase and cell size. Resistance to mitoxantrone conferred only partial resistance to doxorubicin; whereas resistance selected for doxorubicin appeared to confer complete resistance to mitoxantrone. Both agents selected for cross resistance to the Vinca alkaloids. MCF7/D40 cells display a classic-multi-drug resistance phenotype with expression of P-glycoprotein, decreased drug accumulation relative to the parental line and reversal of drug accumulation and drug resistance by verapamil. MCF7/Mitox cells likewise display resistance to multiple drugs, but in contrast to MCF7/D40 cells do not express P-glycoprotein by immunoblot or RNA blot analysis. Net drug accumulation in MCF7/Mitox cells was decreased relative to the parental cells but there was no selective modulation of drug accumulation or in vitro drug resistance by the addition of verapamil. Efflux of mitoxantrone was enhanced in both the MCF7/D40 and MCF7/Mitox cell lines relative to the MCF7/S cell line. We conclude that the two drug resistant cell lines have different mechanisms of decreased drug accumulation.


Relationships between Psychosocial Resilience and Physical Health Status of Western Australian Urban Aboriginal Youth.
Psychosocial processes are implicated as mediators of racial/ethnic health disparities via dysregulation of physiological responses to stress. Our aim was to investigate the extent to which factors previously documented as buffering the impact of high-risk family environments on Aboriginal youths' psychosocial functioning were similarly beneficial for their physical health status. We examined the relationship between psychosocial resilience and physical health of urban Aboriginal youth (12-17 years, n = 677) drawn from a representative survey of Western Australian Aboriginal children and their families. A composite variable of psychosocial resilient status, derived by cross-classifying youth by high/low family risk exposure and normal/abnormal psychosocial functioning, resulted in four groups- Resilient, Less Resilient, Expected Good and Vulnerable. Separate logistic regression modeling for high and low risk exposed youth revealed that Resilient youth were significantly more likely to have lower self-reported asthma symptoms (OR 3.48, p<.001) and carer reported lifetime health problems (OR 1.76, p<.04) than Less Resilient youth. The findings are consistent with biopsychosocial models and provide a more nuanced understanding of the patterns of risks, resources and adaptation that impact on the physical health of Aboriginal youth. The results support the posited biological pathways between chronic stress and physical health, and identify the protective role of social connections impacting not only psychosocial function but also physical health. Using a resilience framework may identify potent protective factors otherwise undetected in aggregated analyses, offering important insights to augment general public health prevention strategies.


Renal outcomes and blood pressure patterns in diabetic and nondiabetic individuals at high cardiovascular risk.
Diabetes and hypertension are risk factors for renal and cardiovascular outcomes. Data on the association of achieved blood pressure (BP) with renal outcomes in patients with and without diabetes are sparse. We investigated the association of achieved SBP, DBP with renal outcomes and urinary albumin excretion (UAE) in people with vascular disease. In this pooled analysis, we assessed renal outcome data from high-risk patients aged 55 years or older with a history of cardiovascular disease, 70% of whom had hypertension, randomized to The Ongoing Telmisartan Alone and in Combination with Ramipril Global Endpoint Trial and to Telmisartan Randomized Assessment Study in ACE Intolerant Subjects with Cardiovascular Disease trials investigating telmisartan, ramipril and their combination with a median follow-up of 56 months. Standardized office BP was measured every 6 months, estimated glomerular filtration rate (eGFR) and UAE at baseline, 2 years and study end. Associations of mean achieved BP on treatment were investigated on major renal outcomes including end-stage renal disease (ESRD), decline of eGFR by at least 40%, doubling of creatinine and the composites thereof and on UAE. Analyses were by Cox regression analysis, analysis of variance and Chi2-test. Of 30 937 patients with complete data, 19 450 patients without and 11 487 with diabetes were enrolled between 1 December 2001 and 31 July 2003 and followed until 31 July 2008. Data were pooled as the outcomes for telmisartan 80 mg/day (n = 2903) or placebo (n = 2907) for Telmisartan Randomized Assessment Study in ACE Intolerant Subjects with Cardiovascular Disease and ramipril 10 mg/day (n = 8407), telmisartan 80 mg/day (n = 8386) or the combination of both (n = 8334) were similar. For both those with and without diabetes, the hazard ratios for the composites ESRD or doubling of serum creatinine (707 events overall) and ESRD or 40% eGFR loss (2371 events overall) reached a nadir at achieved SBP of 120 to less than 140 mmHg, and increased with higher and lower SBP with similar relative risk with or without diabetes. For example, risk for the former composite reached a hazard ratios 3.06 (confidence interval 1.90-4.92) with a mean achieved SBP more than 160 mmHg compared with 120 to less than 130 mmHg with diabetes and hazard ratios 2.14 (1.09-4.26) without diabetes. In contrast, the development of new microalbuminuria and macroalbuminuria (3002 and 846 events overall) associated linearly over the whole range of achieved SBP (apart from a slight increase in risk at SBP less than 120 mmHg only in those without diabetes). Absolute risks for the composite and albuminuria outcomes were consistently greater in those with diabetes as compared with without diabetes with high event rates over the whole SBP spectrum. The increased renal risk at low SBP was not related to a meaningful reduction of mandated study drugs or open label renin-angiotensin-aldosterone system inhibition. In patients at high cardiovascular risk, SBP levels more than 140 mmHg and less than 120 are associated with increased risk for renal outcomes. Renal risk was greater in diabetes across the whole range of achieved SBP and DBP. These data suggest similar target BP range in patients with and without diabetes to prevent renal outcomes, a frequent complication in high-risk vascular patients. Clinical Trial registration: http://clinicaltrials.gov.Unique identifier: NCT00153101.


The effect of Mycobacterium bovis (Bacillus Calmette-Guérin) on macrophage random migration, chemotaxis, and pinocytosis.
Macrophage random migration, chemotactic responsiveness, and pinocytosis were studied in the guinea pig after Bacillus Calmette-Guérin (BCG) administration. Oil-induced peritoneal exudate cells were evaluated for random migration and chemotaxis in modified Boyden chambers. Pinocytosis was measured by the in vitro uptake of colloidal 198Au. There was a parallel increase in random migration and chemotaxis after an i.p. dose of 1.0 X 10(7) COLONY-FORMING UNIts of BCG. This was seen as early as Day 4, sustained through Day 21, and absent by Day 44. A number of different strains and preparations of BCG vaccine exerted a similar augmentation of macrophage mobility. In parallel assays, aliquots of macrophages from BCG-treated animals showed increased pinocytosis, first noted at Day 14, peaking at Day 28, and no longer measurable at Day 44. These alterations in macrophage function following BCG administration may in part contribute to the antitumor effect of BCG adjuvant therapy.


Aldehyde Dehydrogenase Mutation Exacerbated High-Fat-Diet-Induced Nonalcoholic Fatty Liver Disease with Gut Microbiota Remodeling in Male Mice.
Mitochondrial aldehyde dehydrogenase 2 (ALDH2) is a critical enzyme involved in ethanol clearance in acetaldehyde metabolism and plays a key role in protecting the liver. The ALDH2*2 mutation causes a significant decrease in acetaldehyde scavenging capacity, leading to the accumulation of acetaldehyde after consuming alcohol. The prevalence of the ALDH2*2 variant is in 45% of Taiwanese individuals. ALDH2 reportedly has protective properties on myocardial damage, stroke, and diabetic retina damage. However, the effects of ALDH2 in the modulation of metabolic syndromes remain unclear. This study evaluates the roles of ALDH2 in a high-fat-diet-induced metabolic syndrome in mice. Male (M) and female (F) wild-type (WT) and ALDH2 knock-in C57BL/6J mice (4-5 weeks old) were fed a high-fat diet for 16 weeks. Results showed that the body and white-adipose-tissue weights were significantly increased in ALDH2-M compared to those in the other groups. We observed markedly elevated serum levels of alanine transaminase and glucose. Oral glucose-tolerance test and homeostasis-model assessment of insulin resistance (HOMA-IR) values were significantly higher in ALDH2-M mice than those in WT-M mice, with no observable differences in female mice. Abundant steatosis and inflammatory cells were observed in ALDH2-M, with significantly decreased expression of hepatic genes IRS2, GLUT4, and PGC-1α compared to that in WT-M. ALDH2 gene mutation also affected the β-diversity of gut microbiota in ALDH2-M resulting in the decreased abundance of Actinobacteria and an increase in Deferribacteres. Our results suggest that potential changes in gut microbiota may be associated with the defective ALDH2 exacerbation of high-fat-diet-induced liver diseases in male mice. However, female mice were not affected, and sex hormones may be an important factor that requires further investigation.


[Acute left ventricular systolic dysfunction after pericardial effusion drainage].
A patient with a thymoma and initially normal ventricular systolic function developed cardiac tamponade, which was relieved by pericardiocentesis. After four days, the tumor was removed and, one week after the relief of tamponade, she developed severe left ventricular systolic dysfunction, that recovered in three days with venous therapy.


Recent developments in peripheral nerve surgery. Management of open traumatic peripheral nerve lesions.
The modern technique of operative repair of traumatic nerve interruption is discussed. The results of microsurgical repair have proved to be superior to those of conventional methods. Besides the anatomy and the operative technique, the indications and timing for surgical intervention are discussed. Reference to the literature and the authors' own experience show that delayed primary repair is the method of choice.


Single-unit responses to natural vestibular stimuli and eye movements in deep cerebellar nuclei of the alert rhesus monkey.
To study the possible role of the cerebellum in the vestibular-ocular reflex, extracellular responses of cerebellar nuclear neurons were recorded in awake monkeys during natural vestibular stimulation; 115 neurons in the fastigial nucleus responded to horizontal sinusoidal accelerations applied to the head by means of whole-body rotation. More than 75% of these cells were located in a distinct layer, 500 mum thick, in the rostral part of the fastigial nucleus; they were excited by contralateral horizontal angular acceleration and inhibited by ipsilateral rotation (type IIf neurons). The remaining 25% of the population were scattered more caudally in the nucleus, and were excited by ipsilateral rotation and inhibited by contralateral rotation (type If). All showed fairly high resting discharges, averaging 50 spikes/s. Sinusoidal horizontal rotation (0.2--4.8 HZ) produced clear periodic modulation of the firing rate of fastigial neurons, which was approximately sinusoidal about the resting rate at low frequencies. As the frequency of oscillation (and the applied acceleration) increased, the sinusoidal modulation of unit firing increased in amplitude; at high stimulus frequencies the firing rate was usually driven to zero during the inhibitory part of stimulus cycle, but did not saturate in the excitatory half leading to an increase in the mean firing rate. The maximum firing rates of fastigial neurons were related to the peak acceleration by a power function. At all stimulus frequencies, the peak firing frequency of fastigial neurons lagged the input angular acceleration. Maximum firing of most units occurred just prior to the maximum velocity of the head. The gain and phase lag of the averaged unit discharge relative to head acceleration were calculated by Fourier analysis, using the fundamental as a first approximation of the response. Over a 20-fold stimulus range (0.2--4.0 HZ), mean phage lags of type IIf unit responses with respect to the applied acceleration remained relatively constant; the phase lag at 0.9 HZ measured 62 plus or minus 13 degrees. This phase lag is very similar to that recorded from vestibular nerve fibers (15), suggesting that type IIf fastigial neurons provide an excitatory signal to the ipsilateral vestibular nuclei which is in phase with direct vestibular afferent input, although functionally opposite in sign. Over the same frequency range, the gain decreased at minus- 18 dB/decade. Our data suggests that the majority of fastigial neurons work in parallel with flocculus Purkinje cells to functionally inhibit type Iv neurons in the ipsilateral vestibular nuclei.


Relationship between supra-annual trends in influenza rates and stroke occurrence.
Stroke occurrence appears to be a random event, yet annual and supra-annual periodicity is observed. Recent attention in atherosclerotic disease etiology has focused on infectious and inflammatory mechanisms. Influenza is one such infection that may influence stroke occurrence. We explored population-based time series data on stroke occurrence and influenza activity. Using Fourier transformation to isolate low-frequency signals in the data, the inverse transformed time series were regressed using Prais-Winsten regression to correct for serially auto-correlated residuals, to assess the relationship between influenza rates and stroke occurrence rates. Changes in the low-frequency components of influenza activity predicted the changes in low-frequency components of the stroke occurrence data with a delay of about 20 weeks. The delay between changes in influenza activity and subsequent stroke activity was different for different stroke types. Overall, the effect size was small with a tripling of the influenza rate associated with about a 6% change in stroke occurrence rate. A small proportion of the patterns of stroke occurrence may be explained by variation in influenza activity. Further evaluation of influenza as a triggering agent in stroke is needed.


"Self-care selfies": Patient-uploaded videos capture meaningful changes in dexterity over 6 months.
Upper extremity function reflects disease progression in multiple sclerosis (MS). This study evaluated the feasibility, validity, and sensitivity to change of remote dexterity assessments applying human pose estimation to patient-uploaded videos. A discovery cohort of 50 adults with MS recorded "selfie" videos of self-care tasks at home: buttoning, brushing teeth, and eating. Kinematic data were extracted using MediaPipe Hand pose estimation software. Clinical comparison tests were grip and pinch strength, 9 hole peg test (9HPT), and vibration, and patient-reported dexterity assessments (ABILHAND). Feasibility and acceptability were evaluated (Health-ITUES framework). A validation cohort (N = 35) completed 9HPT and videos. The modality was feasible: 88% of the 50 enrolled participants uploaded ≥3 videos, and 74% completed the study. It was also usable: assessments easy to access (95%), platform easy to use (97%), and tasks representative of daily activities (86%). The buttoning task revealed four metrics with strong correlations with 9HPT (nondominant: r = 0.60-0.69, dominant: r = 0.51-0.57, P < 0.05) and ABILHAND (r = -0.48, P = 0.05). Retest validity at 1 week was stable (r > 0.8). Cross-sectional correlations between video metrics and 9HPT were similar at 6 months, and in the validation cohort (nondominant: r = 0.46, dominant: r = 0.45, P < 0.05). Over 6 months, pinch strength (5.8-5.0 kg/cm2 , P = 0.05) and self-reported pinch (ABILHAND) decreased marginally. While only 15% of participants worsened by 20% on 9HPT, 70% worsened in key buttoning video metrics. Patient-uploaded videos represent a novel, patient-centered modality for capturing dexterity that appears valid and sensitive to change, enhancing its potential to be disseminated for neurological disease monitoring and treatment.


Expression analysis of regulatory microRNAs in bovine cumulus oocyte complex and preimplantation embryos.
MicroRNAs (miRNAs) are small endogenous molecules that are involved in a diverse of cellular process. However, little is known about their abundance in bovine oocytes and their surrounding cumulus cells during oocyte development. To elucidate this situation, we investigated the relative expression pattern of sets of miRNAs between bovine oocyte and the surrounding cumulus cells during in vitro maturation using miRNA polymerase chain reaction (PCR) array. Results revealed that a total of 47 and 51 miRNAs were highly abundant in immature and matured oocytes, respectively, compared with their surrounding cumulus cells. Furthermore, expression analysis of six miRNAs enriched in oocyte miR-205, miR-150, miR-122, miR-96, miR-146a and miR-146b-5p at different maturation times showed a dramatic decrease in abundance from 0 h to 22 h of maturation. The expression of the same miRNAs in preimplantation stage embryos was found to be highly abundant in early stages of embryo development and decreased after the 8-cell stage to the blastocyst stage following a typical maternal transcript profile. Similar results were obtained by localization of miR-205 in preimplantation stage embryos, in which signals were higher up to the 4-cell stage and reduced thereafter. miR-205 and miR-210 were localized in situ in ovarian follicles and revealed a spatio-temporal expression during follicular development. Interestingly, the presence or absence of oocytes or cumulus cells during maturation was found to affect the expression of miRNAs in each of the two cell types. Hence, our results showed the presence of distinct sets of miRNAs in oocytes or cumulus cells and the presence of their dynamic degradation during bovine oocyte maturation.


NSAID-activated gene-1 as a molecular target for capsaicin-induced apoptosis through a novel molecular mechanism involving GSK3beta, C/EBPbeta and ATF3.
Capsaicin, a natural product of the Capsicum species of red peppers, is known to induce apoptosis and suppress growth. Non-steroidal anti-inflammatory drug-activated gene-1 (NAG-1) is a cytokine associated with pro-apoptotic and antitumorigenic property in colorectal and lung cancer. Our data demonstrate that capsaicin leads to induction of apoptosis and up-regulates NAG-1 gene expression at the transcriptional level. Overexpression of CCAAT/enhancer binding protein beta (C/EBPbeta) caused a significant increase of basal and capsaicin-induced NAG-1 promoter activity. We subsequently identified C/EBPbeta binding sites in the NAG-1 promoter responsible for capsaicin-induced NAG-1 transactivation. Electrophoretic mobility shift assay and chromatin immunoprecipitation assay confirmed binding of C/EBPbeta to the NAG-1 promoter. Capsaicin treatment resulted in an increase of phosphorylated serine/threonine residues on C/EBPbeta, and the immunoprecipitation study showed that capsaicin enhanced binding of C/EBPbeta with glycogen synthase kinase 3beta (GSK3beta) and activating transcription factor 3 (ATF3). The phosphorylation and interaction of C/EBPbeta with GSK3beta and ATF3 are decreased by the inhibition of the GSK3beta and Protein Kinase C pathways. Knockdown of C/EBPbeta, GSK3beta or ATF3 ameliorates NAG-1 expression induced by capsaicin treatment. These data indicate that C/EBPbeta phosphorylation through GSK3beta may mediate capsaicin-induced expression of NAG-1 and apoptosis through cooperation with ATF3 in human colorectal cancer cells.


Biodefense research and the U.S. regulatory structure whither nonhuman primate moral standing?
Biodefense and emerging infectious disease animal research aims to avoid or ameliorate human disease and suffering arising from the natural outbreak or intentional deployment of some of the world's most dreaded pathogens. Research to develop medical countermeasures to these diseases faces a difficult challenge since the products usually cannot be tested for efficacy in human beings. The U.S. Food and Drug Administration's Animal Rule may be increasingly used to overcome this challenge by allowing researchers to translate animal data into medical countermeasures without human subject efficacy testing. Yet the Animal Rule also has significant implications for increased intensive nonhuman primate research. We argue that despite the common belief that nonhuman primates have a fairly high level of moral standing and the protections for animals that are crucial to the U.S. regulations guiding animal research, the Animal Rule specifically and the regulations generally raise serious problems for the attribution of moral standing to nonhuman primates. We argue, however, that the burden of proof is on a position denying all moral standing to nonhuman primates and compare the implications of the U.S. regulatory structure in this regard with some recent developments in the European Union.


Directional Selection Rather Than Functional Constraints Can Shape the G Matrix in Rapidly Adapting Asexuals.
Genetic covariances represent a combination of pleiotropy and linkage disequilibrium, shaped by the population's history. Observed genetic covariance is most often interpreted in pleiotropic terms. In particular, functional constraints restricting which phenotypes are physically possible can lead to a stable G matrix with high genetic variance in fitness-associated traits, and high pleiotropic negative covariance along the phenotypic curve of constraint. In contrast, population genetic models of relative fitness assume endless adaptation without constraint, through a series of selective sweeps that are well described by recent traveling wave models. We describe the implications of such population genetic models for the G matrix when pleiotropy is excluded by design, such that all covariance comes from linkage disequilibrium. The G matrix is far less stable than has previously been found, fluctuating over the timescale of selective sweeps. However, its orientation is relatively stable, corresponding to high genetic variance in fitness-associated traits and strong negative covariance-the same pattern often interpreted in terms of pleiotropic constraints but caused instead by linkage disequilibrium. We find that different mechanisms drive the instabilities along vs. perpendicular to the fitness gradient. The origin of linkage disequilibrium is not drift, but small amounts of linkage disequilibrium are instead introduced by mutation and then amplified during competing selective sweeps. This illustrates the need to integrate a broader range of population genetic phenomena into quantitative genetics.


A mixed methods analysis of participation in a social contact survey.
Social contact survey data forms a core component of modern epidemic models: however, there has been little assessment of the potential biases in such data. We conducted focus groups with university students who had (n = 13) and had never (n = 14) completed a social contact survey during the COVID-19 pandemic. Qualitative findings were explored quantitatively by analysing participation data. The opportunity to contribute to COVID-19 research, to be heard and feel useful were frequently reported motivators for participating in the contact survey. Reductions in survey engagement following lifting of COVID-19 restrictions may have occurred because the research was perceived to be less critical and/or because the participants were busier and had more contacts. Having a high number of contacts to report, uncertainty around how to report each contact, and concerns around confidentiality were identified as factors leading to inaccurate reporting. Focus groups participants thought that financial incentives or provision of study results would encourage participation. Incentives could improve engagement with social contact surveys. Qualitative research can inform the format, timing, and wording of surveys to optimise completion and accuracy.


Molecular cloning of a novel member of the GLUT family of transporters, SLC2a10 (GLUT10), localized on chromosome 20q13.1: a candidate gene for NIDDM susceptibility.
Non-insulin-dependent diabetes mellitus (NIDDM) is a multifactoral disease with both environmental and genetics causes. Genome-wide screening procedures have identified several susceptibility loci for NIDDM within the human genome. We describe the cloning of a putative sugar transporter that has been localized to human chromosome 20q12-q13.1, one of the genomic loci associated with NIDDM. Because of the strong resemblance of this novel protein to members of the mammalian facilitative glucose transporter family (GLUT), we refer to the protein as GLUT10 (HGMW-approved gene symbol SLC2A10). GLUT10 contains 541 amino acids with several glucose transporter sequence motifs and amino acids essential for glucose transport function. In addition, secondary structure analysis of GLUT10 predicts 12 putative transmembrane domains, a hallmark structure of the GLUT family. The tissue distribution of GLUT10 was determined by Northern analysis, which revealed highest levels of expression in the liver and pancreas. From these data, we believe that the chromosomal localization, tissue distribution, and predicted function make GLUT10 an excellent candidate for a susceptibility gene involved in NIDDM.


Bayesian comparisons of codon substitution models.
In 1994, Muse and Gaut (MG) and Goldman and Yang (GY) proposed evolutionary models that recognize the coding structure of the nucleotide sequences under study, by defining a Markovian substitution process with a state space consisting of the 61 sense codons (assuming the universal genetic code). Several variations and extensions to their models have since been proposed, but no general and flexible framework for contrasting the relative performance of alternative approaches has yet been applied. Here, we compute Bayes factors to evaluate the relative merit of several MG and GY styles of codon substitution models, including recent extensions acknowledging heterogeneous nonsynonymous rates across sites, as well as selective effects inducing uneven amino acid or codon preferences. Our results on three real data sets support a logical model construction following the MG formulation, allowing for a flexible account of global amino acid or codon preferences, while maintaining distinct parameters governing overall nucleotide propensities. Through posterior predictive checks, we highlight the importance of such a parameterization. Altogether, the framework presented here suggests a broad modeling project in the MG style, stressing the importance of combining and contrasting available model formulations and grounding developments in a sound probabilistic paradigm.


Development of moyamoya disease in pregnancy and puerperium: case report.
Progressive moyamoya disease in pregnancy and puerperium has not been reported previously. Here, we present a 39-year-old woman who had been found to have moderate stenosis of right middle cerebral artery (MCA) 4 years prior to her pregnancy, finally suffering minor completed stroke due to progressive moyamoya disease at the early postpartum period. Three days after cesarean section without any complication, she developed cerebral infraction at right hemisphere, when magnetic resonance angiography indicated apparent progression of the proximal MCA stenosis. Catheter angiography demonstrated nearly occlusion of the right terminal internal carotid artery (ICA) and the development of an abnormal vascular network at the base of the brain as well as MCA stenosis, indicating a definitive diagnosis of moyamoya disease with unilateral involvement. The patient underwent superficial temporal artery-middle cerebral artery anastomosis 1 month after the onset of stroke, and she did not manifest as further neurological events during the follow-up period of 2 years. Moyamoya disease could newly develop in pregnancy and puerperium, which should be noted as a pitfall of the management of moyamoya disease with pregnancy.


Effects of natriuretic peptides and nitroprusside on venous function in trout.
Active venous regulation of cardiovascular function is well known in mammals but has not been demonstrated in fish. In the present studies, the natriuretic peptides (NP) rat atrial natriuretic peptide (ANP) and trout ventricular natriuretic peptide (VNP), clearance receptor inhibitor SC-46542, and sodium nitroprusside (SNP) were infused into unanesthetized trout fitted with pressure cannulas in the ventral aorta, dorsal aorta, and ductus Cuvier, and a ventral aorta (VA) flow probe was used to measure cardiac output (CO). In another group, in vivo vascular (venous) capacitance curves were obtained during ANP or SNP infusion. The in vitro effects of NP on vessels and the heart were also examined. ANP, VNP, and SC-46542 decreased central venous pressure (PVen), CO, stroke volume (SV), and gill resistance (RG), whereas systemic resistance (RS) and heart rate (HR) increased. Dorsal aortic pressure (PDA) transiently increased and then fell even though RS remained elevated. ANP decreased mean circulatory filling pressure (MCFP), increased vascular compliance at all blood volumes, and increased unstressed volume in hypovolemic fish. ANP had no direct effect on the heart. ANP responses in vivo were not altered in trout made hypotensive by prior treatment with the angiotensin-converting enzyme inhibitor lisinopril. SNP reduced ventral aortic pressure (PVA), PDA, and RS, increased CO and HR, but did not affect PVen, SV, or RG. SNP slightly decreased MCFP but did not affect compliance or unstressed volume. In vitro, large systemic arteries were more responsive than veins to NP, whereas SNP relaxed both. These results show that, in vivo, NP decrease venous compliance, thereby decreasing venous return, CO, and arterial pressure. Conversely, SNP hypotension is due to decreased RS. This is the first evidence for active regulation of venous capacitance in fish, which probably occurs in small veins or venules. The presence of venous baroreceptors is also suggested.


Comparison of grouper infection with two different iridoviruses using transcriptome sequencing and multiple reference species selection.
Due to high-density aquafarming in Taiwan, groupers are commonly infected with two different iridoviruses: Megalocytivirus (grouper iridovirus of Taiwan, TGIV) and Ranavirus (grouper iridovirus, GIV). Iridoviral diseases cause mass mortality, and surviving fish retain these pathogens, which can then be horizontally transferred. These viruses have therefore become a major challenge for grouper aquaculture. In this study, comparisons of the biological responses of groupers to infection with these two different iridoviruses were performed. A novel approach for transcriptomic analysis was proposed to enhance the discovery of differentially expressed genes and associated biological pathways. In this method, suitable and available reference species are selected from the NCBI taxonomy tree and the Ensembl and KEGG databases instead of either choosing only one model species or adopting the NCBI non-redundant dataset as references. Our results show that selection of multiple appropriate model species as references increases the efficiency and performance of analyses compared to those of traditional approaches. Using this method, 17 shared pathways and 5 specific pathways were found to be significantly differentially expressed following infection with the two iridoviruses, among which 11 pathways were additionally identified based on the proposed method of multiple reference species selection. Among the pathways responsive to infection with a specific iridovirus, the spliceosomal pathway (ko03040; p-value = 0.0011) was exclusively associated with TGIV infection, while the glycolysis/gluconeogenesis pathway (ko00010; p-value = 0.0032) was associated with GIV infection. These findings and designed corresponding biological experiments may facilitate a deeper understanding of the mechanisms by which both TGIV and GIV cause fatal infections, as well as the ways in which they induce different pathologies and symptoms. We believe that the proposed novel mechanism for de novo transcriptomic analysis provides superior and comprehensive functional annotations and that the resulting shared and specific pathways identified may help immunologists develop specific vaccines against various types of iridovirus in the near future.


Acetazolamide attenuates Hunter-Cheyne-Stokes breathing but augments the hypercapnic ventilatory response in patients with heart failure.
Acetazolamide has been used to attenuate Hunter-Cheyne-Stokes breathing with central sleep apnea (CSA) associated with heart failure. However, the mechanisms underlying this improvement remain to be fully elucidated. We hypothesized that acetazolamide stabilizes CSA by attenuating the ventilatory sensitivity to CO2, which is increased in patients with heart failure and is thought to be the major mechanism mediating CSA. Six consecutive male patients with stable systolic heart failure and CSA (apnea-hypopnea index [AHI] ≥ 15 episodes/h) were randomized to a double-blind crossover protocol with acetazolamide or placebo received 1 hour before bedtime for six nights with 2 weeks of wash-out. Under both conditions, we measured the hypercapnic ventilatory response (HCVR), arterial blood Pco2, steady-state metabolic CO2 production, overnight attended polysomnography, and also assessed cardiac and pulmonary function. Compared with placebo, acetazolamide significantly decreased the AHI (65 ± 32 vs. 31 ± 19 events/h, mean ± SD). Acetazolamide increased the HCVR slope by 55% (3.3 ± 1.7 vs. 5.1 ± 2.4 L/min/mm Hg; P = 0.03), an increase that far exceeded the 12% fall in arterial Pco2 (P = 0.02). The acetazolamide-induced change in the balance of these effects (ΔHCVR × Pco2) was inversely associated with the reduction in AHI (r = 0.8; P = 0.045). This placebo-controlled study indicates that acetazolamide improves CSA in patients with heart failure despite an increase in the slope of the HCVR. However, because the degree of HCVR elevation inhibits the improvement in unstable breathing, an increased CO2 chemosensitivity may be a key mechanism underlying an incomplete resolution of CSA with acetazolamide.


Mid- to long-term function and implant survival of ACL reconstruction and medial Oxford UKR.
The purpose of this study was to describe mid- to long-term outcomes of anterior cruciate ligament (ACL) reconstruction with simultaneous or staged medial unicompartmental knee replacement (UKR), and compare outcomes between (1) young patients aged younger than 55 at surgery and those older, (2) those with long-term follow-up greater than 10 years, (3) cemented and cementless UKR, and (4) compare outcomes to those with an intact ACL. We identified knees with staged or simultaneous ACL reconstruction and medial UKR from a prospectively followed designer UKR cohort, and describe mean Oxford Knee Score (OKS), mean Tegner activity score and Kaplan-Meier survival estimates. We matched these knees to ACL-intact knees. Seventy-six consecutive UKR with staged or simultaneous ACL reconstruction were identified with mean six-year follow-up (range 1-15). There was significant improvement in OKS and Tegner score with surgery. At most recent follow-up, OKS was 41.0 (range 11 to 48), and Tegner score 3.6 (0 to 8). There were three revisions occurring at a mean of five years post-operatively. The five-, 10- and 15-year survival estimates were 97% (95% confidence interval [CI] 93-100), 92% (83-100), and 92% (83-100). There was no difference in functional scores or implant survival in young patients, those with long-term follow-up (>10 years), those with cementless fixation, or when compared to ACL intact knees. These results demonstrate excellent mid- to long-term function and survival of selected patients who have undergone ACL reconstruction and medial UKR. Their outcome was similar to those with intact ACLs.


Functional role of the three conserved cysteines in the N domain of visual arrestin-1.
Arrestins specifically bind active and phosphorylated forms of their cognate G protein-coupled receptors, blocking G protein coupling and often redirecting the signaling to alternative pathways. High-affinity receptor binding is accompanied by two major structural changes in arrestin: release of the C-tail and rotation of the two domains relative to each other. The first requires detachment of the arrestin C-tail from the body of the molecule, whereas the second requires disruption of the network of charge-charge interactions at the interdomain interface, termed the polar core. These events can be facilitated by mutations destabilizing the polar core or the anchoring of the C-tail that yield "preactivated" arrestins that bind phosphorylated and unphosphorylated receptors with high affinity. Here we explored the functional role in arrestin activation of the three native cysteines in the N domain, which are conserved in all arrestin subtypes. Using visual arrestin-1 and rhodopsin as a model, we found that substitution of these cysteines with serine, alanine, or valine virtually eliminates the effects of the activating polar core mutations on the binding to unphosphorylated rhodopsin while only slightly reducing the effects of the C-tail mutations. Thus, these three conserved cysteines play a role in the domain rotation but not in the C-tail release.


Sacrificial Silver Nanoparticles: Reducing GeI2 To Form Hollow Germanium Nanoparticles by Electroless Deposition.
Herein we report the electroless deposition of Ge onto sacrificial Ag nanoparticle (NP) templates to form hollow Ge NPs. The formation of AgI is a necessary component for this reaction. Through a systematic study of surface passivating ligands, we determined that tri-n-octylphosphine is necessary to facilitate the formation of hollow Ge NPs by acting as a transport agent for GeI2 and the oxidized Ag(+) cation (i.e., AgI product). Annular dark-field (ADF) scanning transmission electron microscopy (STEM) imaging of incomplete reactions revealed Ag/Ge core/shell NPs; in contrast, completed reactions displayed hollow Ge NPs with pinholes which is consistent with the known method for dissolution of the nanotemplate. Characterization of the hollow Ge NPs was performed by transmission electron microscopy, ADF-STEM, energy-dispersive X-ray spectroscopy, UV-vis spectrophotometry, and Raman spectroscopy. The galvanic replacement reaction of Ag with GeI2 offers a versatile method for controlling the structure of Ge nanomaterials.


Intra-alveolar distraction osteogenesis in preparation for dental implant placement combined with orthodontic/orthognathic surgical treatment: a case report.
The use of implants has become a routine part of restorative dental treatment. The placement of implants requires a sufficient quantity of bone in which to place implant fixtures. Where such bone does not exist, a number of methods have been used to augment the affected area of the alveolar ridge. This case report describes the use of the relatively new distraction osteogenesis procedure to increase bone volume in a patient who required post-traumatic restoration of several missing teeth, as well as comprehensive orthodontic and orthognathic surgical treatment.


Percutaneous needle biopsy of mediastinal masses under C-arm conebeam CT guidance: diagnostic performance and safety.
The aim of this study is to evaluate the feasibility of percutaneous needle biopsy of mediastinal masses under conebeam computed tomography (CBCT) and "XperGuide" navigation guidance. From September 2013 to April 2016, 40 patients (25 men and 15 women; mean age 52.5 years; range 18.7-86.4 years) with 40 mediastinal masses underwent CBCT-/"XperGuide"-guided percutaneous needle biopsies. Technical success, sensibility, specificity, positive predictive value (ppv), negative predictive value (npv) and complications rate were evaluated. Technical success evaluated as the correct positioning of the needle inside the lesion was 100%. Based on histopathological diagnosis, 2 of 40 biopsies (5%) resulted a false negative. Diagnostic accuracy was 95%, sensitivity was 95%, specificity was 100%, ppv was 100%, and npv was 33%. The mean total procedure time was 32 min (range 15-60 min) revealing a reduction in time comparing the first group of 20 patients (mean time 45 min) with the last group of 20 patients (mean time 17 min). No major complications were recorded. Only three patients (7.5%) had mild pneumothorax as demonstrated by post-procedural CBCT, resolved spontaneously. CBCT/"XperGuide" navigation system is a new, safe and accurate technique that can be used as guidance for mediastinal mass biopsies. It also permits the use of CT machines for diagnostic examinations relieving them from interventional procedures burden.


An increased burden of common and rare lipid-associated risk alleles contributes to the phenotypic spectrum of hypertriglyceridemia.
Earlier studies have suggested that a common genetic architecture underlies the clinically heterogeneous polygenic Fredrickson hyperlipoproteinemia (HLP) phenotypes defined by hypertriglyceridemia (HTG). Here, we comprehensively analyzed 504 HLP-HTG patients and 1213 normotriglyceridemic controls and confirmed that a spectrum of common and rare lipid-associated variants underlies this heterogeneity. First, we demonstrated that genetic determinants of plasma lipids and lipoproteins, including common variants associated with plasma triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) from the Global Lipids Genetics Consortium were associated with multiple HLP-HTG phenotypes. Second, we demonstrated that weighted risk scores composed of common TG-associated variants were distinctly increased across all HLP-HTG phenotypes compared with controls; weighted HDL-C and LDL-C risk scores were also increased, although to a less pronounced degree with some HLP-HTG phenotypes. Interestingly, decomposition of HDL-C and LDL-C risk scores revealed that pleiotropic variants (those jointly associated with TG) accounted for the greatest difference in HDL-C and LDL-C risk scores. The APOE E2/E2 genotype was significantly overrepresented in HLP type 3 versus other phenotypes. Finally, rare variants in 4 genes accumulated equally across HLP-HTG phenotypes. HTG susceptibility and phenotypic heterogeneity are both influenced by accumulation of common and rare TG-associated variants.


Evidence for p53 as guardian of the cardiomyocyte mitochondrial genome following acute adriamycin treatment.
The present study is an initial analysis of whether p53 may function as guardian of the cardiomyocyte mitochondrial genome, with mitochondrial p53 localization proposed to be involved in both mitochondrial DNA (mtDNA) repair and apoptosis. Subcellular distribution, protein levels, and possible function(s) of p53 protein in the response of cardiomyocytes to adriamycin (ADR) were analyzed. Levels and subcellular localization of proteins were determined by Western blot and immunogold ultrastructural analysis techniques. Here we demonstrate that stress caused by ADR induced upregulation of p53 protein in cardiomyocyte mitochondria and nuclei between 3 and 24 hr. Increased expression of PUMA and Bax proteins, pro-apoptotic targets of p53, was documented following ADR treatment and was accompanied by increased levels of apoptotic markers, with elevation of cytosolic cytochrome c at 24 hr and subsequent caspase-3 cleavage at 3 days. Mitochondrial p53 levels correlated with mtDNA oxidative damage. Loss of p53 in knockout mouse heart resulted in a significant increase in mtDNA vulnerability to damage following ADR treatment. Our results suggest that mitochondrial p53 could participate in mtDNA repair as a first response to oxidative damage of cardiomyocyte mtDNA and demonstrate an increase of apoptotic markers as a result of mitochondrial/nuclear p53 localization.


Probing lymphocyte biology by genomic-scale gene expression analysis.
The identity and abundance of mRNA species within a cell dictate, to a large extent, the biological potential of that cell. Although posttranscriptional mechanisms modify protein expression in critical ways, cellular differentiation requires key changes in gene transcription, as evidenced by the potent phenotypes that result from disruption of transcription factor genes in mice. It is now possible to assess the mRNA profile of a cell globally using recently developed genomics techniques. This review focuses on the potential of cDNA microarrays to define gene expression in lymphoid cells, a field which is in its infancy. Examples of cellular activation genes and cytokine inducible genes discovered using this technology are presented but these represent only a taste of the fruit that this new technology will ultimately bear. Gene expression profiles should provide essential new insights into lymphocyte differentiation and activation, the pathogenesis of immune disorders, and the molecular abnormalities in lymphoid malignancies.


The MEK inhibitor U0126 ameliorates diabetic cardiomyopathy by restricting XBP1's phosphorylation dependent SUMOylation.
Background: Chronic diabetes accelerates vascular dysfunction often resulting in cardiomyopathy but underlying mechanisms remain unclear. Recent studies have shown that the deregulated unfolded protein response (UPR) dependent on highly conserved IRE1α-spliced X-box- binding protein (XBP1s) and the resulting endoplasmic reticulum stress (ER-Stress) plays a crucial role in the occurrence and development of diabetic cardiomyopathy (DCM). In the present study, we determined whether targeting MAPK/ERK pathway using MEK inhibitor U0126 could ameliorate DCM by regulating IRE1α-XBP1s pathway. Method: Three groups of 8-week-old C57/BL6J mice were studied: one group received saline injection as control (n=8) and two groups were made diabetic by streptozotocin (STZ) (n=10 each). 18 weeks after STZ injection and stable hyperglycemia, one group had saline treatment while the second group was treated with U0126 (1mg/kg/day), 8 weeks later, all groups were sacrificed. Cardiac function/histopathological changes were determined by echocardiogram examination, Millar catheter system, hematoxylin-eosin staining and western blot analysis. H9C2 cardiomyocytes were employed for in vitro studies. Results: Echocardiographic, hemodynamic and histological data showed overt myocardial hypertrophy and worsened cardiac function in diabetic mice. Chronic diabetic milieu enhanced SUMOylation and impaired nuclear translocation of XBP1s. Intriguingly, U0126 treatment significantly ameliorated progression of DCM, and this protective effect was achieved through enriching XBP1s' nuclear accumulation. Mechanistically, U0126 inhibited XBP1s' phosphorylation on S348 and SUMOylation on K276 promoting XBP1s' nuclear translocation. Collectively, these results identify that MEK inhibition restores XBP1s-dependent UPR and protects against diabetes-induced cardiac remodeling. Conclusion: The current study identifies previously unknown function of MEK/ERK pathway in regulation of ER-stress in DCM. U0126 could be a therapeutic target for the treatment of DCM.


Views on contentious practices in dairy farming: the case of early cow-calf separation.
The public has become increasingly interested in the welfare of food animals, but the food animal industries possess few mechanisms for public engagement. Here we present results from a web-based forum designed to allow stakeholders to share views on controversial issues in dairying. In response to the question "Should dairy calves be separated from the cow within the first few hours after birth?" participants were able to indicate "yes," "no," or "neutral" and either write a reason in support of their view or select reasons provided by other participants. Four independent groups of participants were recruited (a total of 163 people); 31% said they had no involvement in the dairy industry; the remaining 69% (with some involvement in the industry) were students or teachers (33%), animal advocates (13%), producers (11%), veterinarians (9%) and other dairy industry professionals (3%). Overall, little consensus existed among participants across groups; 44% chose "yes," 48% "no," and 9% "neutral." Responses varied with demographics, with opposition to early separation higher among females, animal advocates, and those with no involvement with the dairy industry. A fifth group was recruited at a dairy industry conference (an additional 28 participants); 46% chose "yes," 32% "no," and 21% "neutral." Across all 5 groups, opponents and supporters often referenced similar issues in the reasons they provided. Opponents of early separation contended that it is emotionally stressful for the calf and cow, it compromises calf and cow health, it is unnatural, and the industry can and should accommodate cow-calf pairs. In contrast, supporters of early separation reasoned that emotional distress is minimized by separating before bonds develop, that it promotes calf and cow health, and that the industry is limited in its ability to accommodate cow-calf pairs. These results illustrate the potential of web-based forums to identify areas of agreement and conflict among stakeholders, providing a basis for the development of practices that address shared concerns.


Bioactivity and chemical composition of the essential oil from the leaves of Guatteria australis A.St.-Hil.
Essential oil from the leaves of Guatteria australis was obtained by hydrodistillation, analyzed by Gas Chromatography coupled to Mass Spectromery (GC-MS) and their antiproliferative, antileishmanial, antibacterial, antifungal and antioxidant activities were also evaluated. Twenty-three compounds were identified among which germacrene B (50.66%), germacrene D (22.22%) and (E)-caryophyllene (8.99%) were the main compounds. The highest antiproliferative activity was observed against NCI-ADR/RES (TGI = 31.08 μg/ml) and HT-29 (TGI = 32.81 μg/ml) cell lines. It also showed good antileishmanial activity against Leishmania infantum (IC50 = 30.71 μg/ml). On the other hand, the oil exhibited a small effect against Staphylococcus aureus ATCC 6538, S. aureus ATCC 14458 and Escherichia coli ATCC 10799 (MIC = 250 μg/ml), as well as small antioxidant activity (457 μmol TE/g) assessed through ORACFL assay. These results represent the first report regarding chemical composition and bioactivity of G. australis essential oil.


Chronic Losartan Treatment Up-Regulates AT1R and Increases the Heart Vulnerability to Acute Onset of Ischemia and Reperfusion Injury in Male Rats.
Inhibition of angiotensin II type 1 receptor (AT1R) is an important therapy in the management of hypertension, particularly in the immediate post-myocardial infarction period. Yet, the role of AT1R in the acute onset of myocardial ischemia and reperfusion injury still remains controversial. Thus, the present study determined the effects of chronic losartan treatment on heart ischemia and reperfusion injury in rats. Losartan (10 mg/kg/day) was administered to six-month-old male rats via an osmotic pump for 14 days and hearts were then isolated and were subjected to ischemia and reperfusion injury in a Langendorff preparation. Losartan significantly decreased mean arterial blood pressure. However, heart weight, left ventricle to body weight ratio and baseline cardiac function were not significantly altered by the losartan treatment. Of interest, chronic in vivo losartan treatment significantly increased ischemia-induced myocardial injury and decreased post-ischemic recovery of left ventricular function. This was associated with significant increases in AT1R and PKCδ expression in the left ventricle. In contrast, AT2R and PKCε were not altered. Furthermore, losartan treatment significantly increased microRNA (miR)-1, -15b, -92a, -133a, -133b, -210, and -499 expression but decreased miR-21 in the left ventricle. Of importance, addition of losartan to isolated heart preparations blocked the effect of increased ischemic-injury induced by in vivo chronic losartan treatment. The results demonstrate that chronic losartan treatment up-regulates AT1R/PKCδ and alters miR expression patterns in the heart, leading to increased cardiac vulnerability to ischemia and reperfusion injury.


pi-Stacking interactions. Alive and well in proteins.
A representative set of high resolution x-ray crystal structures of nonhomologous proteins have been examined to determine the preferred positions and orientations of noncovalent interactions between the aromatic side chains of the amino acids phenylalanine, tyrosine, histidine, and tryptophan. To study the primary interactions between aromatic amino acids, care has been taken to examine only isolated pairs (dimers) of amino acids because trimers and higher order clusters of aromatic amino acids behave differently than their dimer counterparts. We find that pairs (dimers) of aromatic side chain amino acids preferentially align their respective aromatic rings in an off-centered parallel orientation. Further, we find that this parallel-displaced structure is 0.5-0.75 kcal/mol more stable than a T-shaped structure for phenylalanine interactions and 1 kcal/mol more stable than a T-shaped structure for the full set of aromatic side chain amino acids. This experimentally determined structure and energy difference is consistent with ab initio and molecular mechanics calculations of benzene dimer, however, the results are not in agreement with previously published analyses of aromatic amino acids in proteins. The preferred orientation is referred to as parallel displaced pi-stacking.


Aspirin and dipyridamole in the prevention of acute coronary thrombosis complicating coronary angioplasty.
To test the hypothesis that pretreatment with adequate antiplatelet therapy reduces the likelihood of acute coronary thrombosis during routine percutaneous transluminal coronary angioplasty (PTCA), we reviewed, blinded to treatment group, the films and records of 300 consecutive initially successful PTCAs. Films before PTCA, immediately after, and at least 30 min after the last balloon inflation were assessed for the presence of any thrombus at the PTCA site. We excluded 37 patients who received streptokinase before PTCA or who had 100% occlusion or thrombus on pre-PTCA films. New thrombi were classified as clinically significant (defined as causing 100% occlusion or requiring emergency surgery or streptokinase therapy) or as not significant (not causing an acute problem or requiring intervention). Patients were classified into three groups, based on the type and extent of antiplatelet therapy received. Group 1 (no aspirin, n = 121) consisted of patients who did not receive aspirin either before admission or in hospital before PTCA (with or without dipyridamole). Group 2 (standard treatment, n = 110) received aspirin with or without dipyridamole but did not receive both drugs before admission and in hospital before PTCA. Group 3 (maximal treatment, n = 32) received both aspirin and dipyridamole before admission and in hospital before PTCA. New thrombi were detected at 39 (14.8%) PTCA sites, of which 15 (5.7% of all PTCA sites) were considered clinically significant. Group 1 had the highest incidence of both thrombus (21.5%) and clinically significant thrombus (10.7%). A reduction was seen in group 2 in thrombus (11.8%; p = .07) and in clinically significant thrombus (1.8%; p = .005). Group 3 had no thrombus (p = .001) and no clinically significant thrombus (p = .04). In addition to inadequate pretreatment with antiplatelet therapy, univariate analyses demonstrated several other risk factors for thrombus: higher percent diameter stenosis before PTCA (p less than .008), higher platelet count (p = .013), and current smoking (p = .03). Only higher platelet count (p less than .001) and inadequate pretreatment (p = .001) were associated with clinically significant thrombus. Stepwise logistic regression analysis demonstrated that for thrombus, the lack of effective antiplatelet therapy was the most discriminatory variable, followed by current smoking, higher percent diameter stenosis, and dissection. For clinically significant thrombus, once the lack of pretreatment with effective antiplatelet therapy was considered, no other factors added significant discriminatory information.(ABSTRACT TRUNCATED AT 400 WORDS)


Lifestyle Improvements and Vitamin D Supplementation Play an Important Role in the Prevention of Childhood Diabetes.
This study was to investigate the characteristics of insulin secretion and the 25-hydroxyvitamin D3 (25(OH)D3) levels in children with obesity. A retrospective analysis was conducted among children who underwent health checkups in the pediatric healthcare department of our hospital from January 2018 to January 2021, and they were divided into a normal group and an obese group according to their BMI. The insulin secretion and the 25(OH)D3 levels of the two groups of children were compared. A total of 721 children were included in the study, including 591 in the normal group and 130 in the obese group, with an obesity rate of 18.03%. The blood glucose of the normal group was 4.55 ± 1.75 mmol/L, and the 2 h PG was 7.51 ± 2.11 mmol/L; in the obesity group, they were 6.03 ± 2.16 mmol/L and 8.92 ± 3.24 mmol/L, respectively. The FPG and 2 h PG in the obese group were significantly higher than those in the normal group (all P < 0.05). The incidence of IFG/IGT in the normal group was 5.24% (31/591), and the incidence of DM was 3.71% (22/591); the incidence of IFG/IGT in the obese group was 14.62% (19/130), and the incidence of DM was 13.08% (17/130). The incidences of IFG/IGT and DM in the obese group were significantly higher than those in the normal group (P < 0.05). The FINS of the children in the normal group was 18.46 ± 3.15 μU/mL, and the HOMA-IR was 2.64 ± 0.62; the above indicators in the obese group were 19.11 ± 4.72 μU/mL and 3.01 ± 0.83, respectively. The FINS and HOMA-IR in the obese group were significantly higher than those in the normal group (P < 0.05). The serum 25(OH)D3 level in the normal group was 28.15 ± 5.27 ng/mL, of which 556 cases were normal in 25(OH)D3 and 35 cases were deficient in 25(OH)D3. The serum 25(OH)D3 level in the obese group was 24.35 ± 4.51 ng/mL, of which 112 cases were normal in 25(OH)D3 and 18 cases were deficient in 25(OH)D3. The level of serum 25(OH)D3 in the normal group was significantly higher than that in the normal group, and the ratio of 25(OH)D3 deficiency was significantly lower than that in the normal group (P < 0.05). The blood glucose level of childhood obesity was significantly increased, the incidence of abnormal glucose metabolism and diabetes was significantly increased, and the level of 25(OH) vitamin D3 was significantly decreased. Lifestyle improvements and vitamin D supplementation play an important role in the prevention of childhood diabetes. Because the major causes of childhood obesity are excessive caloric intake and lack of exercise, the most effective and direct measures to prevent obesity are a reasonable lifestyle, reasonable eating habits, and moderate exercise. Although genetics are critical, there is no reliable way to eliminate obesity genes in the human body. Therefore, the role of obesity genes is required to be ultimately eliminated by reduced caloric intake and increased physical activity.


[Confidence of 100 per cent sensitivity and specificity].
A new strategy and four new methods are presented to calculate the limits of the confident interval for an estimate of a proportion equal to 1.0 or 0.0. A current formula which includes 1/(2n) for continuity correction leads to a confident interval which does not include the parameter estimate. Thus, it is proposed: 1) The exclusion of the factor 1/(2n) in that formula leads to correct most of its inconsistencies. The new strategy assumes that the upper limit of a confident interval when the estimates is 100%. The lower limit is calculated by assuming that there is a proportion in the population, from where the sample was taken, such as the probability of getting 100% in the sample is equal to the probability of falling into type I error of current statistics (0.05, 0.01, etc). Three methods are proposed with this strategy. 2) A combinatorial solution based in the knowledge of the number of individuals at whom the test can be applied. 3) A solution based on the binomial distribution. 4) A solution based on the Poisson distribution.


[Flashback following use of cannabis--a review].
The present study tries to find an answer to the question, whether the consumption of cannabis can cause flashbacks, and, if so, under what circumstances they occur. This is of interest for the treatment, prognosis and also from the legal viewpoint. The definition of a flashback is often unclear. Empirical investigations are often not sufficiently comparable, the supplementary data are very often missing, making the evaluation of the results difficult. Self-diagnosis by the consumer is usually unsuitable for estimating the frequency and phenomenology of flashbacks. Cannabis may trigger flashbacks after consuming hallucinogenes; the probability of the occurrence of a flashback seems to increase with the amount of hallucinogenes a person has consumed. A definite correlation between the amount of hashish consumed and the occurrence of flashback does not exist. Flashbacks have also been reported after consuming cannabis alone. However, data vary regarding the frequency of such flashbacks. According to the available data they occur rarely and require a thorough differential diagnostic evaluation in each individual case. As a rule, the occurrence of a flashback may take place in cases where there is an intake of hallucinogenic drugs in the recent case history.


Endoscopic Treatment of Early Cancer of the Colon.
Colorectal cancer is the fourth most common cancer diagnosis worldwide and the second leading cause of cancer death. In the United States, it is estimated that in 2015 there will be 132,700 new cases of colorectal cancer (representing 8.43% of all new cancer cases) and 49,700 deaths. Colonoscopy plays a fundamental role in the prevention and management of colorectal cancer patients and is used for both the diagnosis and treatment of early colorectal cancer and its precursors. Improvements in colonoscopy preparation, new techniques of adenoma detection, and recent progress in endoscopic imaging methods are providing higher-quality results and reducing the incidence and mortality of the disease. Traditionally, colonoscopy has been used to remove precursor lesions. Invasive cancer was treated by surgical resection with or without chemoradiotherapy. During the past decade, endoscopic resection techniques have advanced, and cancers confined to the mucosal and superficial submucosal layers can now be resected via flexible endoscopes. Therefore, it is important to understand the indications and limitations of endoscopic resection, determine whether the cancer can be curatively resected, and assess the risk of lymph node metastasis, which precludes endoscopic treatment.


TLC separation of hippuric, mandelic, and phenylglyoxylic acids from urine after mixed exposure to toluene and styrene.
A method using thin-layer chromatography is described to determine the concentration of hippuric acid, mandelic acid, and phenylglyoxylic acid present in the urine after occupational mixed exposure to toluene and styrene. These substances are known metabolites of toluene and styrene, and therefore the evaluation to mixed exposure to toluene and styrene may be carried out separating these metabolites beforehand. Procedures are proposed to separate the metabolites as follows: (1) separation of hippuric acid from mandelic acid, (2) separation of mandelic acid from phenylglyoxylic acid, and (3) separation of hippuric acid and mandelic acid from phenylglyoxylic acid. The developing reagent p-dimethylaminobenzaldehyde in acetic acid anhydride was used after separation on Kieselgel and Silicagel. The sensitivity of the method was 6 microgram of hippuric acid, 10 microgram of mandelic acid, and 7 microgram of phenylglyoxylic acid with an average recovery of 94%.


Effects of different oocyte retrieval and in vitro maturation systems on bovine embryo development and quality.
Cyclic adenosine monophosphate (cAMP) modulators have been used to avoid spontaneous oocyte maturation and concomitantly improve oocyte developmental competence. The current work evaluated the effects of the addition of cAMP modulators forskolin, 3-isobutyl-1-methylxanthine (IBMX) and cilostamide during in vitro maturation on the quality and yields of blastocysts. The following experimental groups were evaluated: (i) slicing or (ii) aspiration and maturation in tissue culture medium (TCM)199 for 24 h (TCM24slicing and TCM24aspiration, respectively), (iii) aspiration and maturation in the presence of cAMP modulators for 30 h (cAMP30aspiration) and in vivo-produced blastocysts. In vitro-matured oocytes were fertilized and presumptive zygotes were cultured in vitro to assess embryo development. Cleavage, blastocyst formation, blastocyst cell number, mRNA abundance of selected genes and global methylation profiles were evaluated. Blastocyst rate/zygotes for the TCM24aspiration protocol was improved (32.2 ± 2.1%) compared with TCM24slicing and cAMP30aspiration (23.4 ± 1.2% and 23.3 ± 2.0%, respectively, P 0.05), while those from the other groups were significantly elevated. It is concluded that retrieval, collection systems and addition of cAMP modulators can affect oocyte developmental competence, which is reflected not only in blastocyst rates but also in global DNA methylation and gene expression patterns.


Cost-effectiveness of statins.
Currently, 6 hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are marketed in the United States (US). Given the wide variation in the prices and efficacy of statins, formal cost-effectiveness analysis may improve drug selection decisions. To assess the cost-effectiveness of statin therapy in primary and secondary prevention of coronary heart disease, we developed a model of the costs and consequences of lipid-regulating therapy and estimated the incremental cost-effectiveness of 5 statins (atorvastatin, fluvastatin, lovastatin, pravastatin, simvastatin) at usual starting doses versus no therapy. Drug effects on serum lipids were assessed using data approved by the US Food and Drug Administration for product labeling. Annual risks of coronary event occurrence were estimated using Framingham Heart Study coronary risk equations developed for use in this model. Current estimates of direct medical costs of coronary heart disease were used to assign costs to health states and acute coronary events. Main outcome measurements were net cost (statin therapy minus savings in coronary heart disease treatment), gain in life expectancy, and cost per life-year saved. The maximum gain in life expectancy was achieved with atorvastatin, which also had a lower net cost than lovastatin, pravastatin, and simvastatin. Compared with fluvastatin, atorvastatin's greater effectiveness is attained at a lower cost per life-year saved. The cost-effectiveness of HMG-CoA reductase inhibition in primary and secondary prevention of coronary heart disease has been improved with the introduction of atorvastatin.


Vaccine hesitancy among parents of adolescents and its association with vaccine uptake.
Addressing parental vaccine hesitancy may increase adolescent vaccination acceptance. However, no validated measure exists to identify parents hesitant toward adolescent vaccines. To determine if a modified version of the Parent Attitudes about Childhood Vaccines (PACV) survey, a previously validated tool to identify parental hesitancy toward vaccines in infants, predicts adolescent vaccine uptake at office visits. We modified the PACV for use in the adolescent setting and distributed it to a convenience sample of parents of adolescents aged 11 to 17 presenting for care at a diverse group of six pediatric practices in Oklahoma and South Carolina. We determined the vaccination status of the parents' adolescents for 3 vaccines (Tetanus-diphtheria-acellular pertussis [Tdap], meningococcal conjugate [MCV4], and human papillomavirus [HPV] vaccines). We used Fisher's exact tests to compare vaccination status with each survey item and with an overall general hesitancy scale that we constructed. We analyzed 363 surveys. At the time of the visit, vaccination coverage was 84% for Tdap, 73% for MCV, and 45% for any dose of HPV. Thirty-nine percent of parents expressed concern about vaccine efficacy and 41% expressed concern about side effects. Forty-five percent of parents disagreed with the statement that "teens can get all of the vaccines that are due at a single visit." Two individual items were associated with not receiving a dose of HPV vaccine that was due. The overall modified PACV score failed to predict adolescent vaccine uptake at an office visit. Several individual items were associated with vaccine uptake. The cumulative modified PACV, a general measure of vaccine hesitancy, was not associated with vaccination status despite illuminating parental hesitancy. We need to better understand vaccine-specific concerns for the adolescent population.


Angiotensin-converting enzyme determination in plasma during therapy with converting enzyme inhibitor: two methods compared.
For normal and above-normal concentrations of angiotensin-converting enzyme (ACE; EC 22.214.171.124) activity in plasma, results of a manual fluorometric method [with hippuryl-histidyl-leucine (HHL), 5 mmol/L, as substrate] correlated well with those of an automated spectrophotometric method [with 3-(2-furylacryloyl)-L-phenylalanyl-glycyl-glycine (FAPGG), 2 mmol/L, as substrate]. However, for patients receiving converting enzyme inhibitor (CEI) therapy, the spectrophotometric method showed much greater suppression of plasma ACE activity than did the fluorometric method. To determine which of the two methods provided a more reliable indication of ACE inhibition in vivo, we measured plasma ACE, angiotensin I (ANG I), and angiotensin II (ANG II) in patients receiving the CEI perindopril. During perindopril therapy, changes in the ratio of ANG II:ANG I, an index of ACE activity in vivo, showed a close agreement with changes in plasma ACE activity measured with FAPGG as substrate, but not with HHL as substrate. We conclude that measurement of ACE activity in vitro with FAPGG as substrate provides a reliable measure of changes in conversion of ANG I to ANG II in vivo during CEI therapy.


Clinical evaluation of nedocromil sodium in asthma.
The recognition that inflammatory events in the airways play a key role in the pathogenesis of asthma has led to a relentless search for pharmacological agents which modify these processes. Nedocromil sodium (Tilade) represents one such agent. Nedocromil sodium, when inhaled by patients with asthma (0.05-0.50% nebulized, 0.5-4.0 mg m.d.i.), has been shown to inhibit immediate bronchoconstriction provoked by challenges with allergen (10 studies), exercise (five studies), isocapnic hyperventilation, fog and sulphur dioxide (one study each) and adenosine (two studies). With these challenges, inhibition of bronchoconstriction exhibited dose-dependency up to 4 mg, with nedocromil sodium being up to four times more potent than sodium cromoglycate. When inhaled prior to allergen provocation, nedocromil sodium inhibited the late asthmatic reaction; when taken regularly during the pollen season, it attenuated the allergen-induced increase in non-specific bronchial responsiveness. The efficacy of nedocromil sodium (4 mg q.i.d.) in the treatment of clinical asthma was initially shown in four open studies and subsequently confirmed in nine double-blind, placebo-controlled 4-12 week studies on patients with seasonal and perennial asthma. Further clinical trials (eight studies) identified some difficulty in replacing inhaled corticosteroids with nedocromil sodium, especially if the corticosteroids were reduced rapidly (four studies). However, two studies have shown that nedocromil sodium produced further improvement in asthma symptoms when used in addition to bronchodilators and inhaled corticosteroids. Treatment with nedocromil sodium (4 mg q.i.d.) for up to 52 weeks demonstrated a progressive reduction in bronchodilator usage throughout the whole treatment period. During clinical assessment, nedocromil sodium was well tolerated, side-effects being unpleasant taste, nausea and headache. In most cases the adverse reactions were mild and transient, although in approximately 3% of patients they resulted in withdrawal from clinical trials. Thus, nedocromil sodium is a novel drug of proven efficacy in the treatment of asthma. Its position in the therapeutic armamentarium is likely to be as an adjunct to bronchodilators and inhaled steroids, to produce improvement in symptoms beyond that achieved with the already established drugs.


Effectiveness of hyaluronan on corneal epithelial barrier function in dry eye.
The aim of this study was to assess quantitatively the effectiveness of hyaluronan on corneal disruption in patients with dry eye. Corneal epithelial barrier function was evaluated by measuring fluorescein permeability using a slit-lamp fluorophotometer. 11 patients with dry eye were assigned to this study. Hyaluronan ophthalmic solution (0.1% hyaluronic acid) was instilled five times a day to the right eye, in addition to the usual artificial tear solutions. The left eye received only the artificial tear solutions. Corneal barrier function was evaluated on the pretreatment day, and at 2 and 4 weeks after treatment. Fluorophotometry was used to measure fluorescein uptake at the central and lower corneal portions. Two weeks after treatment, hyaluronan treated right corneas showed significant corneal epithelial barrier improvement in the lower portion, compared with the pretreatment day (p < 0.025). Four weeks after treatment, the treated corneas showed significant improvement in the central corneal portion (p < 0.025) and improvement in the lower portion, compared with the pretreatment day. The untreated left corneas, on the other hand showed no improvement during the course of the study. This study suggests that hyaluronan is effective in the treatment of corneal epithelial disruption in dry eye.