The Metal Drives the Chemistry: Dual Functions of Acireductone Dioxygenase.
Acireductone dioxygenase (ARD) from the methionine salvage pathway (MSP) is a unique enzyme that exhibits dual chemistry determined solely by the identity of the divalent transition-metal ion (Fe2+ or Ni2+) in the active site. The Fe2+-containing isozyme catalyzes the on-pathway reaction using substrates 1,2-dihydroxy-3-keto-5-methylthiopent-1-ene (acireductone) and dioxygen to generate formate and the ketoacid precursor of methionine, 2-keto-4-methylthiobutyrate, whereas the Ni2+-containing isozyme catalyzes an off-pathway shunt with the same substrates, generating methylthiopropionate, carbon monoxide, and formate. The dual chemistry of ARD was originally discovered in the bacterium Klebsiella oxytoca, but it has recently been shown that mammalian ARD enzymes (mouse and human) are also capable of catalyzing metal-dependent dual chemistry in vitro. This is particularly interesting, since carbon monoxide, one of the products of off-pathway reaction, has been identified as an antiapoptotic molecule in mammals. In addition, several biochemical and genetic studies have indicated an inhibitory role of human ARD in cancer. This comprehensive review describes the biochemical and structural characterization of the ARD family, the proposed experimental and theoretical approaches to establishing mechanisms for the dual chemistry, insights into the mechanism based on comparison with structurally and functionally similar enzymes, and the applications of this research to the field of artificial metalloenzymes and synthetic biology.


A Case of Advanced Unicentric Retroperitoneal Castleman's Disease, Associated With Psoriasis.
We present here a 32-year-old male with advanced lately diagnosed, right sided retroperitoneal mass, which had been already treated due to progressive muco-cutaneous lesions clinically consistent with psoriasis, during recent four years. The advanced retroperitoneal mass resected surgically and reported as hyaline-vascular castleman disease with a dense focus of coarse calcification, on histopathology. Association of psoriasis and castlman disease is discussed in this case report.


Secondhand smoke exposure of children at home and prevalence of parental smoking following implementation of the new tobacco control law in Macao.
To investigate secondhand smoke exposure (SHS) of children at home and the prevalence of parental smoking after implementation of the new tobacco control law in Macao. This study explored whether the smoking ban in public places in Macao has decreased the prevalence of smoking or led to increased SHS exposure of children at home. As smokers cannot smoke in public places any more, they may smoke at home more frequently; a displacement effect of smoke-free legislation. Cross-sectional survey. This study surveyed 337 fathers and 538 mothers. Questions from a subset of key questions from the Global Adult Tobacco Survey (2nd edition) were applied to assess the SHS exposure of children and the prevalence of parental smoking since the smoking ban. A classification tree analysis was used to analyse the factors increasing SHS exposure of children. The prevalence of SHS exposure in children at home was 41.3%. The prevalence rates of paternal and maternal smoking were 43.7% and 3.8%, respectively. Compared with data reported by the Health Bureau of Macao SAR in 2011, the prevalence of parental smoking and the prevalence of SHS exposure of children at home have not decreased since the smoking ban. Analysis of the factors increasing the prevalence of SHS exposure of children indicated that fathers with an education level below high school were more likely to contribute to this increase, compared with fathers with a high school education or more (48.2% vs 32.4%, respectively). In addition, fathers represented the majority of smokers at home, accounting for 92.0% of 415 smoking parents. The prevalence of paternal smoking (82.0%) in the group of children with SHS exposure was much higher than that in the unexposed group (16.7%, Chi-squared test = 367.199, P = 0.000). The SHS exposure of children increased consistently with the decrease in paternal education level. This was consistent with the increasing prevalence of paternal smoking as paternal education level decreased. SHS exposure was most common among children whose fathers had an education level below high school and whose mothers were aged ≤29 years (75.0%). This study did not find any decline in the prevalence of parental smoking after the smoking ban. These parents were more likely to smoke at home after the ban, leading to more frequent SHS exposure for their children.


Disrupted Control Network Connectivity in Abstinent Patients with Alcohol Dependence.
Alcohol causes damage to the brain and is associated with various functional impairments. However, much of the brain damage can be reversed by abstaining for enough time. This study aims to investigate the patterns and degrees of brain function in abstinent patients with alcohol dependence by using resting-state functional connectivity. 26 male patients with alcohol dependence (alcohol group) and 28 age-matched male healthy volunteers (control group) were recruited from a mental hospital and the community, respectively. Using 3T MRI scan data, the resting-state functional connectivity of the task-negative and task-positive networks was determined and compared between the groups. There were no significant group differences in the resting-state functional connectivity in the default mode or in the salience and sensorimotor networks. Compared with the control group, the alcohol group showed significantly lower functional connectivity in the executive control network, especially in the cingulo-opercular network and, in some regions of interest, the dorsal attention network. This finding suggests that some brain networks do not normalize their functions after abstinence from drinking, and these results may be helpful in future research to investigate the mechanisms for craving alcohol and alcohol relapse prevention.


The protocol of tagging endogenous proteins with fluorescent tags using CRISPR-Cas9 genome editing.
The currently widely used CRISPR-Cas9 genome editing technology enables the editing of target genes (knock-out or knock-in) with high accuracy and efficiency. Guided by the small guide RNA, the Cas9 nuclease induces a DNA double-strand break at the targeted genomic locus. The DNA double-strand break can be repaired by the homology-directed repair pathway in the presence of a repair template. With the repair template containing the coding sequence of a fluorescent tag, the targeted gene can be inserted with the sequence of a fluorescent tag at the designed position. The genome editing mediated labeling of endogenous proteins with fluorescent tags avoids the potential artifacts caused by gene overexpression and substantially improves the reproductivity of imaging experiments. This protocol focuses on creating mammalian cell lines with endogenous proteins tagged with fluorescent proteins or self-labeling protein tags using CRISPR-Cas9 genome editing.


Codon Usage and Phenotypic Divergences of SARS-CoV-2 Genes.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which first occurred in Wuhan (China) in December of 2019, causes a severe acute respiratory illness with a high mortality rate, and has spread around the world. To gain an understanding of the evolution of the newly emerging SARS-CoV-2, we herein analyzed the codon usage pattern of SARS-CoV-2. For this purpose, we compared the codon usage of SARS-CoV-2 with that of other viruses belonging to the subfamily of Orthocoronavirinae. We found that SARS-CoV-2 has a high AU content that strongly influences its codon usage, which appears to be better adapted to the human host. We also studied the evolutionary pressures that influence the codon usage of five conserved coronavirus genes encoding the viral replicase, spike, envelope, membrane and nucleocapsid proteins. We found different patterns of both mutational bias and natural selection that affect the codon usage of these genes. Moreover, we show here that the two integral membrane proteins (matrix and envelope) tend to evolve slowly by accumulating nucleotide mutations on their corresponding genes. Conversely, genes encoding nucleocapsid (N), viral replicase and spike proteins (S), although they are regarded as are important targets for the development of vaccines and antiviral drugs, tend to evolve faster in comparison to the two genes mentioned above. Overall, our results suggest that the higher divergence observed for the latter three genes could represent a significant barrier in the development of antiviral therapeutics against SARS-CoV-2.


A Multitarget Therapeutic Peptide Derived From Cytokine Receptors Based on in Silico Analysis Alleviates Cytokine-Stimulated Inflammation.
Septicemia is a severe inflammatory response caused by the invasion of foreign pathogens. Severe sepsis-induced shock and multiple organ failure are the two main causes of patient death. The overexpression of many proinflammatory cytokines, such as TNF-α, IL-1β, and IL-6, is closely related to severe sepsis. Although the treatment of sepsis has been subject to many major breakthroughs of late, the treatment of patients with septic shock is still accompanied by a high mortality rate. In our previous research, we used computer simulations to design the multifunctional peptide KCF18 that can bind to TNF-α, IL-1β, and IL-6 based on the binding regions of receptors and proinflammatory cytokines. In this study, proinflammatory cytokines were used to stimulate human monocytes to trigger an inflammatory response, and the anti-inflammatory ability of the multifunctional KCF18 peptide was further investigated. Cell experiments demonstrated that KCF18 significantly reduced the binding of proinflammatory cytokines to their cognate receptors and inhibited the mRNA and protein expressions of TNF-α, IL-1β, and IL-6. It could also reduce the expression of reactive oxygen species induced by cytokines in human monocytes. KCF18 could effectively decrease the p65 nucleus translocation induced by cytokines, and a mice endotoxemia experiment demonstrated that KCF18 could reduce the expression of IL-6 and the increase of white blood cells in the blood stimulated by lipopolysaccharides. According to our study of tissue sections, KCF18 alleviated liver inflammation. By reducing the release of cytokines in plasma and directly affecting vascular cells, KCF18 is believed to significantly reduce the risk of vascular inflammation.


Effect of hypertension on angiotensin-(1-7) levels in rats with different angiotensin-I converting enzyme polymorphism.
To determine circulating angiotensin-(1-7) [Ang-(1,7)] levels in rats with different angiotensin converting enzyme (ACE) genotypes and to evaluate the effect of hypertension on levels of this heptapeptide, plasma levels of angiotensin II (Ang II) and Ang-(1-7) were determined by HPLC and radioimmunoassay in (a) normotensive F0 and F2 homozygous Brown Norway (BN; with high ACE) or Lewis (with low ACE) rats and (b) in hypertensive F2 homozygous male rats (Goldblatt model). Genotypes were characterized by PCR and plasma ACE activity measured by fluorimetry. Plasma ACE activity was 2-fold higher (p < 0.05) in homozygous BN compared to homozygous Lewis groups. In the Goldblatt groups, a similar degree of hypertension and left ventricular hypertrophy was observed in rats with both genotypes. Plasma Ang II levels were between 300-400% higher (p < 0.05) in the BN than in the Lewis rats, without increment in the hypertensive animals. Plasma Ang-(1-7) levels were 75-87% lower in the BN rats (p < 0.05) and they were significantly higher (p < 0.05) in the hypertensive rats from both genotypes. Plasma levels of Ang II and Ang-(1-7) levels were inversely correlated in the normotensive rats (r = -0.64; p < 0.001), but not in the hypertensive animals. We conclude that there is an inverse relationship between circulating levels of Ang II and Ang-(1-7) in rats determined by the ACE gene polymorphism. This inverse relation is due to genetically determined higher ACE activity. Besides, plasma levels of Ang-(1-7) increase in renovascular hypertension.


Long term respiratory sequelae of whooping cough in a nationally representative sample.
The long term respiratory consequences of whooping cough in childhood were sought among members of the National Survey of Health and Development. Peak expiratory flow rate was measured when the survey members were 36 years old and seven respiratory symptoms were reported at the same time. Peak expiratory flow rate was slightly reduced in those who had had whooping cough as a child, and this difference became non-significant when other factors were taken into account. Unexpectedly, chronic cough was significantly less likely to be reported by those who had had whooping cough, and this difference remained significant only among men after other factors had been taken into account. This study failed to show a long term detrimental effect of whooping cough on the respiratory system.


Quality of life in long-stay institutions in England: nurse and resident perceptions.
Quality of life (QOL) of older persons living in institutions is a critical consideration in international economic policies and health care decisions. Yet, there is little relevant research to support decisions about culturally acceptable and effective nursing care for this population. This qualitative study explored and compared perceptions of older residents and nurses in three long-stay institutions in south-west England. It addressed issues viewed as important to a good QOL for residents and ways in which nurses promote QOL. A convenience sample of 10 residents aged 65 and older (80% female) and nine nurses (n = five registered general nurses and four enrolled nurses) were interviewed using a semi-structured schedule. Interview data were analysed using an analytic induction method of content analysis. While theme frequency and examples differed, themes common to both residents and nurses were individuality, professionalism, connectedness, and physical functioning. Resident responses tended to be personal, succinct, and sometimes negative. Nurses' responses were more positive, detailed, and reflective of general professional responsibilities. Implications for nursing education, research, and practice are discussed. Improvement in QOL will be addressed differently based on social and cultural settings. However, there may be commonalities that are applicable across cultures.


Testing chemical carcinogenicity by using a transcriptomics HepaRG-based model?
The EU FP6 project carcinoGENOMICS explored the combination of toxicogenomics and in vitro cell culture models for identifying organotypical genotoxic- and non-genotoxic carcinogen-specific gene signatures. Here the performance of its gene classifier, derived from exposure of metabolically competent human HepaRG cells to prototypical non-carcinogens (10 compounds) and hepatocarcinogens (20 compounds), is reported. Analysis of the data at the gene and the pathway level by using independent biostatistical approaches showed a distinct separation of genotoxic from non-genotoxic hepatocarcinogens and non-carcinogens (up to 88 % correct prediction). The most characteristic pathway responding to genotoxic exposure was DNA damage. Interlaboratory reproducibility was assessed by blindly testing of three compounds, from the set of 30 compounds, by three independent laboratories. Subsequent classification of these compounds resulted in correct prediction of the genotoxicants. As expected, results on the non-genotoxic carcinogens and the non-carcinogens were less predictive. In conclusion, the combination of transcriptomics with the HepaRG in vitro cell model provides a potential weight of evidence approach for the evaluation of the genotoxic potential of chemical substances.


Improving risk prediction for depression via Elastic Net regression - Results from Korea National Health Insurance Services Data.
Depression, despite its high prevalence, remains severely under-diagnosed across the healthcare system. This demands the development of data-driven approaches that can help screen patients who are at a high risk of depression. In this work, we develop depression risk prediction models that incorporate disease co-morbidities using logistic regression with Elastic Net. Using data from the one million twelve-year longitudinal cohort from Korean National Health Insurance Services (KNHIS), our model achieved an Area Under the Curve (AUC) of the Receiver Operating Characteristic (ROC) of 0.7818, compared to a traditional logistic regression model without co-morbidity analysis (AUC of 0.6992). We also showed co-morbidity adjusted Odds Ratios (ORs), which may be more accurate independent estimate of each predictor variable. In conclusion, inclusion of co-morbidity analysis improved the performance of depression risk prediction models.


[Modified total pelvic floor reconstruction for repair of severe pelvic organ prolapse].
To evaluate the clinical outcome of a novel approach for pelvic floor reconstruction using synthetic mesh (modified total pelvic floor reconstruction) for treatment of pelvic organ prolapse. Modified total pelvic floor reconstruction was performed in 30 patients with severe pelvic organ prolapse (including vault prolapse). The clinical outcome of each patient was assessed. The mean operation time was (74.2 +/- 21.5) minutes, and the mean blood loss was (103.3 +/- 40.1) ml. Among them, 23 patients (76.7%) were able to micturate spontaneously the next morning after surgery, with residual urine less than 100 ml. The mean post-operative hospital stay was (4.2 +/- 1.8) days. All patients were followed up for a medium of 6 months. Totally 93.3% and 96.7% of patients were objectively (according to Pelvic Organ Prolapse Quantitive Examination score) and subjectively (according to Prolapse Quality of Life) cured, respectively. During follow-up, only one patient was found to have asymptomatic erosion, and de novo urgent urinary incontinence was seen in 6.7% of patients. The most prominent complication was dyspareunia (66.7%). Although the post-operative sexual function was reported to be worse, no significant difference between patients' pre- and post-operative Pelvic Organ Prolapse/ Urinary Incontinence Sexual Function Questionnaire-12 scores was noted. Modified total pelvic floor reconstruction is a safe, effective, and micro-invasive approach for severe pelvic organ prolapse repair; however, dyspareunia remains a main concern.


Symptom dimensions in OCD: item-level factor analysis and heritability estimates.
To reduce the phenotypic heterogeneity of obsessive-compulsive disorder (OCD) for genetic, clinical and translational studies, numerous factor analyses of the Yale-Brown Obsessive Compulsive Scale checklist (YBOCS-CL) have been conducted. Results of these analyses have been inconsistent, likely as a consequence of small sample sizes and variable methodologies. Furthermore, data concerning the heritability of the factors are limited. Item and category-level factor analyses of YBOCS-CL items from 1224 OCD subjects were followed by heritability analyses in 52 OCD-affected multigenerational families. Item-level analyses indicated that a five factor model: (1) taboo, (2) contamination/cleaning, (3) doubts, (4) superstitions/rituals, and (5) symmetry/hoarding provided the best fit, followed by a one-factor solution. All 5 factors as well as the one-factor solution were found to be heritable. Bivariate analyses indicated that the taboo and doubts factor, and the contamination and symmetry/hoarding factor share genetic influences. Contamination and symmetry/hoarding show shared genetic variance with symptom severity. Nearly all factors showed shared environmental variance with each other and with symptom severity. These results support the utility of both OCD diagnosis and symptom dimensions in genetic research and clinical contexts. Both shared and unique genetic influences underlie susceptibility to OCD and its symptom dimensions.


Rationale for angiotensin II receptor blocker therapy in chronic heart failure.
Inhibiting the synthesis of angiotensin II (Ang II) by angiotensin-converting enzyme (ACE) inhibitor treatment has long been established as the recommended therapy for heart failure (HF). As ACE inhibitors are only partially effective in blocking the formation of Ang II, additional treatment with selective Ang II receptor blockers (ARBs) promise additional benefits in patients with HF. This hypothesis is currently being evaluated in Val-HeFT, a large-scale, randomised, double-blind, placebo-controlled study on the effects of adding the highly selective ARB valsartan to usual therapy (diuretics, digoxin, beta-blockers, ACE inhibitors) for HF treatment.


Time-resolved pH/pO2 mapping with luminescent hybrid sensors.
A method for simultaneous and referenced 2D mapping of pH and pO2 is described. The experimental setup combines a fast gateable CCD camera as detector, a LED as excitation light source and a single-layer sensor membrane as optical transducer. The planar optode comprises a lipophilic fluorescein derivative (lifetime approximately 5 ns) and platinum(II) mesotetrakis(pentafluorophenyl)porphyrin (approximately 70 micros in the absence of a quencher) immobilized in a hydrogel matrix. Depending on the fluorescent pH indicator, a pH transition in the physiological range (pH 6-pH 8) or in the near-basic region (pH 7-pH 9) can be achieved. The measuring scheme involves the time-resolved acquisition of images in three windows during a series of square-shaped excitation pulses. A method allowing the calculation of both parameters from these three images is presented. The pH/pO2 hybrid sensor incorporating the pH indicator 2',7'-dihexyl-5(6)-N-octadecyl-carboxamidofluorescein was characterized in detail. The pH and pO2 were determined with a maximum deviation of 0.03 pH unit and 6.5 hPa pO2, respectively, within the range of pH 7.6-pH 8.7 and 0-200 hPa pO2 in test measurements. The ionic strength (IS) cross-sensitivity was found to be relatively small (pH/IS < 3.5 x 10(-4) mM(-1) and pO2/IS <-0.053 hPa mM(-1) at a transition from 0.5 to 0.1 M IS). Whereas a strong temperature effect on the sensor signal was observed (DeltapH/DeltaT = 0.011-0.034 K-1 and DeltapO2/DeltaT = 1.85-7.17 hPa K-1 in the range from 277 to 308 K). Examples of pH/pO2 images obtained in natural marine sediment are presented.


Stiffness Matters: Part I-The Effects of Plate Stiffness on the Biomechanics of ACDF In Vitro.
In vitro biomechanical testing of human cadaveric cervical and goat cervical motion segments. The aim of this study was to measure the effects of plate stiffness on load-sharing, instantaneous axis of rotation (IAR), and posterior element loading after anterior cervical discectomy and fusion (ACDF). ACDF is intended to create an environment, which facilitates sufficient stability and biomechanical conditions to promote bone formation. The relationship between cervical plate stiffness, load-sharing, and the IAR is complex. The ideal cervical plate is sufficiently stiff to limit interbody motion but is compliant enough to facilitate load-sharing rather than stress-shielding. Anterior cervical plates of distinct bending stiffnesses were applied to human and goat cervical motion segments following ACDF. A validated custom force-sensing interbody implant was placed in the disc space to measure load-sharing in the spine. Interbody loads, posterior element strain, and the IAR were measured during flexion/extension for each plate. Load-sharing in the interbody space, posterior element strain, and the location of the IAR were all significantly affected by plate stiffness. More compliant plates resulted in more load sharing, less posterior element strain, and a more dorsally located IAR relative to stiffer plates. A more compliant plate fosters more consistent load-sharing through the entire range of flexion/extension, which may promote faster bone formation and better fusion. A more compliant plate causes less posterior element strain, which may reduce facet joint loads and in turn reduce facet joint arthrosis. An ideal plate may be one that is stiff enough to minimize interbody motion and yet compliant enough to allow consistent load-sharing and minimal increase in posterior element strain. N/A.


Benefit of cardiopoietic mesenchymal stem cell therapy on left ventricular remodelling: results from the Congestive Heart Failure Cardiopoietic Regenerative Therapy (CHART-1) study.
Left ventricular (LV) reverse remodelling is an important marker of improved outcomes in patients with advanced heart failure (HF). We examined the impact of the intramyocardial administration of bone-marrow-derived, lineage-directed, autologous cardiopoietic mesenchymal stem cells (C3BS-CQR-1) on LV remodelling in patients with advanced HF enrolled in the CHART-1 study. Patients (n=351) with symptomatic advanced HF secondary to ischaemic heart disease, and reduced LV ejection fraction (LVEF <35%) were randomized to receive C3BS-CQR-1 or a sham procedure. In a post hoc analysis we examined the effect of C3BS-CQR-1 on LV reverse remodelling within 1 year of the procedure and the influence of C3BS-CQR-1 dosing in the 271 patients treated as randomized. Delivery of C3BS-CQR-1 was associated with a progressive decrease in both LV end-diastolic volume (LVEDV) and end-systolic volume (LVESV) within 52 weeks after treatment. At 1 year, the LVEDV and LVESV of treated patients decreased by 17.0 mL and 12.8 mL greater than controls (P=0.006 and P=0.017, respectively). The effect on LVEDV was maintained after multivariable adjustment for baseline age, systolic blood pressure, LVEDV, LVEF and history of myocardial infarction. The largest reverse remodelling was evident in the patients receiving a moderate number of injections (<20). In CHART-1, intramyocardial administration of cardiopoietic stem cells led to reverse remodelling as evidenced by significant progressive decreases in LVEDV and LVESV through the 52 weeks of follow-up. Further studies are needed to explore the dose response with regard to cell number and injected volume, and reverse remodelling.


Variety and portion size combine to increase food intake at single-course and multi-course meals.
Increases in food variety and portion size independently promote intake. Little is known about how these effects combine or how they depend on meal structure. In two randomized crossover experiments, once a week for four weeks, women ate a lunch meal that was varied in two properties: variety (low: three bowls of the favorite dish vs. high: three bowls, each with a different main dish) and portion size (small: 450 g vs. large: 600 g). In Experiment 1 (n = 42), dishes were served simultaneously and in Experiment 2 (n = 49), dishes were served sequentially over three courses. At each meal, the primary outcome of food intake was measured; additionally, we measured sensory-specific satiety (SSS; the relative hedonic decline of a food as it is eaten). In Experiment 1 (simultaneous structure) variety and portion size did not interact (p = 0.72) but both independently increased intake; participants consumed 15 ± 7 g more at meals with high variety compared to low and 57 ± 7 g more from large portions compared to small (both p < 0.03). Similarly, in Experiment 2 (sequential structure) variety and portion size did not interact (p = 0.99) but participants consumed 30 ± 8 g more at high-variety meals and 51 ± 8 g more from large portions (both p < 0.001). SSS was not influenced by portion size in either experiment (both p > 0.16) or by variety in Experiment 1 (p = 0.58), but SSS was smaller at high-variety meals in Experiment 2 (p = 0.001). Thus, variety and large portions promoted greater food intake for a similar or smaller hedonic decline, indicating these effects were facilitated by delayed SSS. At meals with either a simultaneous or sequential structure, high variety and large portions combined to increase intake, suggesting that these common properties act together to promote overconsumption.


Vascular effects of deletion of melanocortin-4 receptors in rats.
Obesity is a major cause of hypertension, but links between the obese and hypertensive states remain incompletely understood. A major component of cardiovascular function in obese individuals is a state of sympathoactivation. A postulated mechanism of this sympathoactivation is the activation of specific classes of neurons commonly associated with metabolic control, which also affect sympathetic outflow to cardiovascular targets. One class of neurons is characterized by expression of melanocortin-4 receptors (MC4R) which are activated by metabolic signals such as leptin and insulin. In this study, we examined the effects of deletion of MC4R in a novel rat model. MC4R knockout (KO) rats are obese and profoundly insulin resistant without frank diabetes. Despite these conditions, MC4R KO rats are normotensive. Moderate bradycardia and significant increases in peripheral resistance were evident in MC4R KO rats. To determine if the dissociation between hypertension and obesity was associated with changes in vascular function, in vitro reactivity to vasoactive agents and in vivo reactivity to sympathetic blockade were examined. Vasodilator function was not affected by obesity in MC4R KO rats. Reactivity to phenylephrine was reduced, suggesting desensitization of adrenergic signaling. In response to ganglionic blockade with mecamylamine, blood pressure and hindlimb resistance fell more in MC4R KO rats, suggesting that sympathoactivation of the vascular was still evident, despite the absence of hypertension. These findings suggest that obesity causes sympathoactivation of the vasculature despite the absence of MC4R. Dissociation of obesity from hypertension in this model may reflect more renal mechanisms of blood pressure control.


Dietary intake of potassium and associated dietary factors among representative samples of Japanese general population: NIPPON DATA 80/90.
The purpose of this study was to investigate the dietary potassium intake and associated other dietary factors among a representative sample cohort of Japanese population. We obtained data from NIPPON DATA80 and 90 that were conducted with the National Nutrition Surveys in 1980 and in 1990. Then we estimated nutrient and food intakes of individuals in the National Nutrition Survey of 1980 and that of 1990, which were adjusted on the basis of data of the National Nutrition Survey of 1995. We analyzed data for 10,422 participants (4585 men and 5837 women) in NIPPON DATA80 and 8342 participants (3488 men and 4854 women) in NIPPON DATA90 having dietary potassium intake information. In NIPPON DATA80 and 90 it was observed that there was a significant relationship between the dietary potassium intake and age for both men and women. Higher potassium intake was associated with higher age, intake of protein, iron, calcium, sodium, vitamins, and fiber. Regarding food groups, lower amount of dietary cereals, rice, flour, fats and oils were associated with higher dietary potassium for both men and women. On the other hand, higher intake of nuts, potatoes, soy beans, fruits, vegetables, mushrooms, sea algae, fish and shellfish were associated with higher dietary potassium. We obtained the mean dietary potassium intake and its association with other dietary nutrient intake in Japanese adults as the baseline data in NIPPON DATA80 and in NIPPON DATA90.


Impact of Operative Time on Adverse Events Following Primary Total Joint Arthroplasty.
Little is known regarding the impact of operative time on adverse events following arthroplasty. The present study tests for associations between a 15-minute increase in operative time and the occurrence of adverse events following primary total joint arthroplasty. Patients undergoing primary total hip or knee arthroplasty during 2006-2013 were identified in the American College of Surgeons National Surgical Quality Improvement Program. Operative time (as a continuous variable) was tested for association with perioperative outcomes using multivariate regression. All regressions were adjusted for differences in demographic, comorbidity, and procedural characteristics. A total of 165,474 patients met inclusion criteria. The mean (±standard deviation) operative time was 91.9 ± 32.5 minutes. Following adjustment for baseline characteristics, an increase in operative time by 15 minutes increased the risk of anemia requiring transfusion by 9% (95% confidence interval [CI] = 8%-10%, P < .001), wound dehiscence by 13% (95% CI = 8%-19%, P < .001), renal insufficiency by 9% (95% CI = 3%-14%, P < .001), sepsis by 10% (95% CI = 6%-14%, P < .001), surgical site infection by 9% (95% CI = 7%-12%, P < .001), and urinary tract infection by 4% (95% CI = 2%-6%, P < .001). Similarly, an increase in operative time by 15 minutes increased the risk of hospital readmission by 5% (95% CI = 4%-6%, P < .001) and of extended hospital length of stay (≥4 days) by 9% (95% CI = 8%-10%, P < .001). The present study suggests that greater operative time increases the risk for multiple postoperative complications following total joint arthroplasty. These data suggest that surgeons should consider steps to minimize operative time without compromising the technical components of the surgical procedure.


Methylation of HIN-1, RASSF1A, RIL and CDH13 in breast cancer is associated with clinical characteristics, but only RASSF1A methylation is associated with outcome.
Aberrant promoter CpG island hypermethylation is associated with transcriptional silencing. Tumor suppressor genes are the key targets of hypermethylation in breast cancer and therefore may lead to malignancy by deregulation of cell growth and division. Our previous pilot study with pairs of malignant and normal breast tissues identified correlated methylation of two pairs of genes - HIN-1/RASSFIA and RIL/CDH13 - with expression of estrogen receptors (ER), progesterone receptors (PR), and HER2 (HER2). To determine the impact of methylation on clinical outcome, we have conducted a larger study with breast cancers for which time to first recurrence and overall survival are known. Tumors from 193 patients with early stage breast cancer who received no adjuvant systemic therapy were used to analyze methylation levels of RIL, HIN-1, RASSF1A and CDH13 genes for associations with known predictive and prognostic factors and for impact on time to first recurrence and overall survival. In this study, we found that ER was associated with RASSF1A methylation (p < 0.001) and HIN-1 methylation (p = 0.002). PR was associated with RIL methylation (p = 0.012), HIN-1 (p = 0.002), and RASSF1A methylation (p = 0.019). Tumor size was associated with RIL and CDH13 methylation (both p = 0.002), and S-phase was associated with RIL methylation (p = 0.036). Only RASSF1A was associated with worse time to first recurrence (p = 0.045) and worse overall survival (p = 0.016) after adjusting for age, tumor size, S-phase, estrogen receptor and progesterone receptor. Methylation of HIN-1, RASSF1A, RIL and CDH13 in breast cancers was associated with clinical characteristics, but only RASSF1A methylation was associated with time to first recurrence and overall survival. Our data suggest that RASSF1A methylation could be a potential prognostic biomarker.


Ophthalmic fluorometholone-gentamicin versus ophthalmic betamethasone-gentamicin following cataract surgery.
A prospective, randomised, investigator-masked, parallel-group study was performed to compare fluorometholone-gentamicin eye drops and ointment with betamethasone-gentamicin eyedrops and ointment in the control of ocular inflammation after cataract surgery. Seventy patients (35 in each treatment group) of both sexes undergoing cataract-lens implant surgery for visually disabling cataract were enrolled in the study. The demographic and baseline parameters on day I, the day after surgery, were similar in the two study groups. After treatment, on day 3 and day 6 post-operatively, the reduction in cells in the anterior chamber and conjunctival hyperaemia were similar in the two study groups. Both treatments were equally well-tolerated. Ophthalmic fluorometholone-gentamicin was as effective as ophthalmic betamethasone-gentamicin in the control of ocular inflammation after cataract surgery.


[Underestimated frequency of resistant hypertension in predialysis nondiabetic patients].
The real prevalence of resistant hypertension (RH) is unknown. Studies suggest that it affects 10%-15% of patients treated for hypertension by primary care physicians. RH is defined as blood pressure (BP) remaining above the goal despite the use of optimal doses of 3 or more medicines of different classes (including a diuretic). It means BP >140/90 mm Hg for the general population and >130/80 mm Hg for patients with diabetes or kidney disease. Prior to diagnosing a patient as having RH, it is important to document medication compliance and exclude white-coat hypertension, inaccurate BP measurement, and secondary causes. The role of aldosterone in RH has gained increasing recognition. There is strong evidence for the use of spironolactone as a highly effective antihypertensive agent. Aldosterone plays a significant role in RH pathogenesis, primarily due to its vasoconstrictive effects and the possibility of altering vascular compliance. In RH, there is a high prevalence of cardiac and extra-cardiac target organ damage. It is known that BP control in chronic kidney disease is the key factor for reducing cardiovascular risk and renal disease progression. The objective of the study was to evaluate the prevalence of RH in predialysis nondiabetic (CKD stage I-IV) patients.


Generating intrinsically disordered protein conformational ensembles from a Markov chain.
Intrinsically disordered proteins (IDPs) sample a diverse conformational space. They are important to signaling and regulatory pathways in cells. An entropy penalty must be payed when an IDP becomes ordered upon interaction with another protein or a ligand. Thus, the degree of conformational disorder of an IDP is of interest. We create a dichotomic Markov model that can explore entropic features of an IDP. The Markov condition introduces local (neighbor residues in a protein sequence) rotamer dependences that arise from van der Waals and other chemical constraints. A protein sequence of length N is characterized by its (information) entropy and mutual information, MIMC, the latter providing a measure of the dependence among the random variables describing the rotamer probabilities of the residues that comprise the sequence. For a Markov chain, the MIMC is proportional to the pair mutual information MI which depends on the singlet and pair probabilities of neighbor residue rotamer sampling. All 2N sequence states are generated, along with their probabilities, and contrasted with the probabilities under the assumption of independent residues. An efficient method to generate realizations of the chain is also provided. The chain entropy, MIMC, and state probabilities provide the ingredients to distinguish different scenarios using the terminologies: MoRF (molecular recognition feature), not-MoRF, and not-IDP. A MoRF corresponds to large entropy and large MIMC (strong dependence among the residues' rotamer sampling), a not-MoRF corresponds to large entropy but small MIMC, and not-IDP corresponds to low entropy irrespective of the MIMC. We show that MorFs are most appropriate as descriptors of IDPs. They provide a reasonable number of high-population states that reflect the dependences between neighbor residues, thus classifying them as IDPs, yet without very large entropy that might lead to a too high entropy penalty.


A novel polymer-paclitaxel conjugate based on amphiphilic triblock copolymer.
A novel amphiphilic polymer-paclitaxel conjugate P(LGG-paclitaxel)-PEG-P(LGG-paclitaxel) has been prepared. It was derived from its parent polymer P(LGG)-PEG-P(LGG), poly{(lactic acid)-co-[(glycolic acid)-alt-(l-glutamic acid)]}-block-poly(ethylene glycol)-block-poly{(lactic acid)-co-[(glycolic acid)-alt-(l-glutamic acid)]}, which was prepared by ring-opening copolymerization of l-lactide (LLA) with (3s)-benzoxylcarbonylethyl-morpholine-2,5-dione (BEMD) in the presence of dihydroxyl PEG with molecular weight of 4600 as a macroinitiator using stannous octoate (Sn(Oct)(2)) as catalyst, and by subsequent catalytic hydrogenation. It could self-assemble into micelles in an aqueous system with P(LGG-paclitaxel) block in the core and PEG in the shell. ESEM and DLS analysis of the micelles revealed a homogeneous spherical morphology and a unimodal size distribution. In vitro release of paclitaxel from the conjugate micelles showed that its release rate depended on pH value and was higher at lower pH than in neutral condition. In vitro antitumor activity of the paclitaxel conjugate against rat brain glioma C6 cells was evaluated by MTT method. The results showed that the paclitaxel can be released from the conjugate without losing cytotoxicity.


Optical breakdown in transparent media with adjustable axial length and location.
We demonstrate a highly elongated (aspect ratio over 500:1) optical breakdown in water produced by a single pulse of a picosecond laser focused with a combination of an axicon and a lens. Locations of the proximal and distal ends of the breakdown region can be adjusted by modifying radial intensity distribution of the incident beam with an amplitude mask. Using Fresnel diffraction theory we derive a transmission profile of the amplitude mask to create a uniform axial intensity distribution in the breakdown zone. Experimentally observed dynamics of the bubbles obtained with the designed mask is in agreement with the theoretical model. A system producing an adjustable cylindrical breakdown can be applied to fast linear or planar dissection of transparent materials. It might be useful for ophthalmic surgical applications including cataract surgery and crystalline lens softening.


[Hospital financing systems: an analysis of trends and a study of the consistency of reform proposals for introduction into public health systems].
The author comments in this paper on the recent literature on reforms in hospital reimbursement systems. Instead of taking a theoretical approach to this topic, the article adopts a "middle of the road" view, by looking at the consistency of applying some of these reforms (mainly on contract management and DRGs) in Public Health Systems. After recognising some of the existing problems in the management of public hospitals in Spain, the author advocates for a common sense strategy to health reforms in terms of the following: (i) at the time of implementing changes in the hospital sector keep targets defined as simple as possible, (ii) go to reforms through a step-by-step procedure, and (iii) whenever things look complex, use basic "house-keeping" economics, instead of just importing additional economic literature from abroad.


Assessing the severity of depressive states in recently detoxified alcoholics.
The problem of how to assess "unbiased" symptoms of depression meaningfully in the context of alcoholism bedevils studies of concomitant alcoholism and affective disorder. This study summarizes this controversy and discusses its implications for developing improved measures of depression severity among alcoholics. The Beck Depression Inventory (BDI) responses of 130 alcoholic applicants applying for inpatient care were evaluated using a two-parameter normal item response model. This study demonstrates that a single dimension of depression severity accounts for patient responses well, but that seven BDI items were relatively poor markers of syndrome severity for these alcoholics. The study documents growing consensus among investigators as to which BDI items constitute a fair scale of depression severity among alcoholic patients The availability of "unbiased criteria" for assessing the severity of depression among alcoholics applying for inpatient treatment will enable investigators and clinicians to recognize patients with concomitant alcoholism and affective disorder for special attention and/or treatment. The modified BDI scale proposed here is a candidate for a clinical definition of depression severity among alcoholic patients by virtue of this study's demonstration that it defines a distinct syndrome in this patient sample; but the modified scale must undergo independent tests of validity before its clinical utility can be established.


The application of electromagnetic theory to electrocardiology. II. Numerical solution of the integral equations.
In an earlier paper exact integral equations were derived for the surface potentials resulting from sources within an irregularly shaped inhomogeneous body. These exact equations cannot usually be solved. In this paper a discrete analogue is constructed which is not straightforward to solve, but which can be treated by careful mathematical methods. In particular a deflation procedure greatly facilitates the iterative solution of the problem and overcomes the divergence encountered by other authors. Numerical solutions obtained for simple geometries are compared to the exact analytic solutions available in such cases. The necessary convergence of the solutions of the discrete analog towards the solution of the continuous problem is shown to occur only if the coefficients of the discrete analogue are carefully evaluated. Calculations are then presented for realistic thoracic geometries, typical results being presented as surface potential maps. Finally the important effect of the internal regional inhomogeneities, particularly a realistic cardiac blood mass, is demonstrated by obtaining vector loops with and without these effects.


Distribution of Human Papillomavirus Genotypes in Iranian Women According to the Severity of the Cervical Lesion.
Persistent infection with high-risk human papillomavirus (HPV) has been recognized as a major cause of cervical cancer. Distribution of HPV genotypes may differ according to the geographic region and the severity of the cervical lesion. Determining HPV genotypes' specific distribution is useful for HPV surveillance and control programs. However, little is known about the distribution of HPV genotypes in Iranian women. The aim of this study was to determine the distribution of HPV genotypes in Iranian women with different grades of cervical lesions. From 2011 to 2013, a total of 436 Iranian women with convenience sampling strategy were included in this cross-sectional study. In detail, 287 women negative for intraepithelial lesion or malignancy, 32 with atypical squamous cells of undetermined significance (ASCUS), 50 with low-grade squamous intraepithelial lesion (LSIL), 44 with high-grade squamous intraepithelial lesion (HSIL), and 23 with cervical cancer were evaluated in this investigation. HPV genotypes were determined by INNO-LiPA HPV Genotyping Extra assay. In total, HPV infection was detected in 45.4% of the cases. The most common high-risk HPV (HR-HPV) genotype was HPV-16 (32.8%), followed by HPV-53 (9.1%). Within low-risk (LR-HPV) genotypes HPV-6 (22.2%) and HPV-44 (6.1%) were the most prevalent. HPV-16 was the predominant genotype in cases with cervical cancer (56.5%), ASCUS (34.4%), and HSIL (34.1%). HPV-6 was the most common genotype in normal cases (9.1%) and LSIL patients (18%). The prevalence of HPV positivity was significantly higher in cases with high-grade lesions (≥ HSIL) (64.2%) than in normal/LSIL (37.3%) (P = 0.033). The rate of HR-HPV infection was significantly higher in ≥ HSIL cases (61.2%) than normal/LSIL (27.9%) (P = 0.003). This study describes robust information on the distribution of HPV genotypes among Iranian women with and without cervical lesions. The present data may be of importance for designing future public health strategies, including HPV vaccination programs.


Rescue of fertility in homozygous mice for the urokinase plasminogen activator transgene by the transplantation of mouse hepatocytes.
Development of the urokinase plasminogen activator/SCID (uPA/SCID) transgenic mouse model has opened new perspectives for the study of different biological mechanisms such as liver regeneration, stem cell differentiation, and human hepatic pathogens. We observed that homozygous uPA/SCID mice (uPA+/+/SCID) had a small offspring, indicating a fertility defect. The goal of this study was thus to rescue the fertility of homozygous uPA mice. A deregulation of ovarian function with an absence of corpus luteum was observed in female uPA+/+/SCID mice. In male uPA+/+/SCID mice, a decrease of the weight of the testes, epididymis, seminal vesicle, and prostate was measured. This was associated with an absence of seminal and prostatic secretions and a reduction in testicular sperm production. We hypothesized that the infertility of mice was the consequence of uPA-induced liver injury. Thus, in order to rescue liver function, hepatocytes from mice negative for the uPA transgene were transplanted into uPA+/+/SCID mice. Thirty days after cell transplantation, the livers of transplanted uPA+/+/SCID mice were totally repopulated and presented a normal morphology. Furthermore, transplantation restored normal body weight, life span, and reproductive organ function. In conclusion, we demonstrated that the transplantation of uPA+/+/SCID mice with healthy hepatocytes was sufficient to rescue the reproductive capacity of female and male uPA homozygous animals, highlighting the importance of normal liver function to reproductive capability.


2-Amino-alpha-2'-deoxyadenosine increased duplex stability of methoxyethylphosphoramidate alpha-oligodeoxynucleotides with RNA target.
A new efficient synthesis of 2-amino-alpha-2'-deoxyadenosine and its incorporation into methoxyethylphosphoramidate alpha-oligodeoxynucleotides (ODNs) via H-phosphonate chemistry were reported. Thermal denaturation experiments demonstrated a significant stabilization of the complexes formed between these analogues and their RNA target (+2 degrees C/NH2A) relative to adenosine-containing phosphoramidate alpha-oligonucleotides. Concerning the binding specificity of these modified ODNs, unlike natural ODNs, discrimination against G pairing is higher and against C pairing is lower.


Escaping Malthus: Economic Growth and Fertility Change in the Developing World.
Following mid-twentieth century predictions of Malthusian catastrophe, fertility in the developing world more than halved, while living standards more than doubled. We analyze how fertility change related to economic growth during this episode, using data on 2.3 million women from 255 household surveys. We find different responses to fluctuations and long-run growth, both heterogeneous over the life cycle. Fertility was procyclical but declined and delayed with long-run growth; fluctuations late (but not early) in the reproductive period affected lifetime fertility. The results are consistent with models of the escape from the Malthusian trap, extended with a life cycle and liquidity constraints.


Structure of Tetrahymena telomerase reveals previously unknown subunits, functions, and interactions.
Telomerase helps maintain telomeres by processive synthesis of telomere repeat DNA at their 3'-ends, using an integral telomerase RNA (TER) and telomerase reverse transcriptase (TERT). We report the cryo-electron microscopy structure of Tetrahymena telomerase at ~9 angstrom resolution. In addition to seven known holoenzyme proteins, we identify two additional proteins that form a complex (TEB) with single-stranded telomere DNA-binding protein Teb1, paralogous to heterotrimeric replication protein A (RPA). The p75-p45-p19 subcomplex is identified as another RPA-related complex, CST (CTC1-STN1-TEN1). This study reveals the paths of TER in the TERT-TER-p65 catalytic core and single-stranded DNA exit; extensive subunit interactions of the TERT essential N-terminal domain, p50, and TEB; and other subunit identities and structures, including p19 and p45C crystal structures. Our findings provide structural and mechanistic insights into telomerase holoenzyme function.


State-of-art review of current malleable penile prosthesis devices in the commercial market.
The malleable penile implant is often considered an inferior device to the three-piece inflatable penile prosthesis implant. Nonetheless, the malleable prosthesis has its unique advantages such as lower cost, easier to perform and fewer mechanical complications than inflatable prostheses. Furthermore, its role can be extended to patients with issues relating to poor manual hand dexterity, those undergoing a salvage for infection prosthesis and as an emergency surgical measure in patients presenting with acute ischaemic priapism. Over the past few decades, there have been numerous design and technological advancements to improve overall clinical efficacy, mechanical durability, axial rigidity and device concealability of malleable penile prostheses. The following article provides a narrative review of the six major contemporary malleable penile prosthesis devices in the commercial market, namely, the Coloplast Genesis prosthesis, the Boston Scientific Tactra prosthesis, the Zephyr ZSI 100 and 100 (female-to-male) FTM devices, the Rigi10 prosthesis, the TUBE malleable prosthesis and the Shah prosthesis and evaluates the published outcomes. Appropriate patient selection and strict counselling regarding what to expect with malleable prostheses coupled with adherence to safe surgical principles are paramount to ensure excellent clinical success and patient satisfaction rates.


NMR-based lipoprotein analysis for patients with severe hypercholesterolemia undergoing lipoprotein apheresis or PCSK9-inhibitor therapy (NAPALI-Study).
Taking into account the discordance between low-density lipoprotein cholesterol (LDL-C) and LDL particle (LDL-P) number, cardiovascular risk more closely correlates with LDL-P in patients. The aim of our study was to evaluate the number of lipid particles in patients with severe hypercholesterolemia treated with different lipid-lowering regimens. Four groups of patients differing with respect to lipid-lowering therapy were recruited from hypercholesterolemic outpatients and lipoprotein apheresis (LA) facilities, and were treated with statins alone (group A), with statins and proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors (PCSK9i) (group B), with statins and LA (group C), or with statins, PCSK9i, and LA (group D). Cholesterol, triglycerides, LDL-C, high-density lipoprotein cholesterol (HDL-C), LDL-P number and size, HDL-P number and size were determined using nuclear magnetic resonance spectroscopy. The lowest LDL-P number was achieved at the end of LA sessions in combination with statins or in combination with statins and a monoclonal PCSK9i (median; 25th and 75th percentile) (group C: 244 nmoL/L: 237, 244, P < 0.05; group D: 244 nmoL/L: 99, 307, P < 0.05). Comparing LDL-P number at the start of LA (group C: 978 nmoL/L: 728, 1404; group D: 954 nmoL/L: 677, 1521) to the other patient groups (groups A and B), the lowest LDL-P number was measured in patients treated with PCSK9i and a statin (group B): LDL-P (762 nmoL/L: 604, 1043, P < 0.05), large LDL-P (472 nmoL/L: 296, 574, P < 0.05), and small LDL-P (342 nmoL/L: 152, 494, P < 0.05). Very low-density lipoprotein and HDL particle sizes remained approximately the same in all groups. LA in combination with statins or in combination with statins and PCSK9i most reduced LDL-P numbers in hypercholesterolemic patients.


"Living in the moment" among cancer survivors who report life-transforming change.
Some cancer survivors say that while coping with their illness they have had what they identify as a highly beneficial, personal, life-transforming change (LTC; a self-subscribed condition among participants in this study, not a researcher-defined construct). Previously we found that the content of LTC was highly individual; but certain features of the LTC process were quite uniform. For example, LTC was driven by a very rational desire to remain genuinely functional as a person in the midst of highly stressful circumstances. Our purpose is to further understand LTC by analysis of participants' critique of questionnaire items on positive subjective outcomes. We obtained a set of 67 positive psychosocial questionnaire items from the Psychosocial Impact of Illness (PII) item bank development team of the National Institutes of Health (NIH) Patient-Reported Outcomes Measurement Information Systems (PROMIS) initiative. The PII items are sourced from a variety of instruments, representing more than one theory of personal growth. This enabled a grounded theory approach, allowing a theory of LTC to emerge from the data. Each of nine cancer survivors participated in a single, two-part session: an hour-long semi-structured qualitative interview regarding their LTC (previously reported), then individually critiquing the 67 items in terms of relevance and importance to their self-reported LTC experience (their critique is analyzed in this article). The PII items were grouped into 20 researcher-defined conceptual themes and average scores for each theme were computed and compared. The theme "living in the moment" (LITM; sometimes referred to as "Being in the moment" in other literature) scored significantly higher than other themes. Qualitative interview transcripts revealed that participants spontaneously used LITM as a coping tactic that reduced worries to manageable levels. These participants noticed subsequently that LITM also produced unexpected substantial benefits in many aspects of their lives unrelated to cancer. This experience motivated increased use of LITM that led to LTC beyond coping with cancer. LITM can be a practical, effective coping tactic to manage worries and promote adaptation to the challenges of a life-threatening illness. Participant interviews indicated the motivation for LITM is not traumatic experience, but seems to be conservation of their own personal resources for coping with increased stresses. Hobfoll's conservation of resources model appears to be more consistent with LITM among our participants than Tedeshi's posttraumatic growth (PTG) model. Several PII items that represent LITM have unfortunately been removed from the current PROMIS PII item bank, indicating a need for improved of item selection criteria. Further research is needed first to identify or develop assessment items that more effectively capture the precursors and phases of LITM, and then develop interventions that facilitate LITM and LTC for patients.


The effectiveness of position- and composition-specific gap costs for protein similarity searches.
The flexibility in gap cost enjoyed by hidden Markov models (HMMs) is expected to afford them better retrieval accuracy than position-specific scoring matrices (PSSMs). We attempt to quantify the effect of more general gap parameters by separately examining the influence of position- and composition-specific gap scores, as well as by comparing the retrieval accuracy of the PSSMs constructed using an iterative procedure to that of the HMMs provided by Pfam and SUPERFAMILY, curated ensembles of multiple alignments. We found that position-specific gap penalties have an advantage over uniform gap costs. We did not explore optimizing distinct uniform gap costs for each query. For Pfam, PSSMs iteratively constructed from seeds based on HMM consensus sequences perform equivalently to HMMs that were adjusted to have constant gap transition probabilities, albeit with much greater variance. We observed no effect of composition-specific gap costs on retrieval performance. These results suggest possible improvements to the PSI-BLAST protein database search program. The scripts for performing evaluations are available upon request from the authors.


Ultra-High-Throughput Clinical Proteomics Reveals Classifiers of COVID-19 Infection.
The COVID-19 pandemic is an unprecedented global challenge, and point-of-care diagnostic classifiers are urgently required. Here, we present a platform for ultra-high-throughput serum and plasma proteomics that builds on ISO13485 standardization to facilitate simple implementation in regulated clinical laboratories. Our low-cost workflow handles up to 180 samples per day, enables high precision quantification, and reduces batch effects for large-scale and longitudinal studies. We use our platform on samples collected from a cohort of early hospitalized cases of the SARS-CoV-2 pandemic and identify 27 potential biomarkers that are differentially expressed depending on the WHO severity grade of COVID-19. They include complement factors, the coagulation system, inflammation modulators, and pro-inflammatory factors upstream and downstream of interleukin 6. All protocols and software for implementing our approach are freely available. In total, this work supports the development of routine proteomic assays to aid clinical decision making and generate hypotheses about potential COVID-19 therapeutic targets.


Meiotic chromosome synapsis-promoting proteins antagonize the anti-crossover activity of sgs1.
Sgs1, the budding yeast homolog of the mammalian BLM helicase, has been implicated in preventing excess recombination during both vegetative growth and meiosis. Most meiotic crossover (CO) recombination requires full function of a set of yeast proteins (Zip1, Zip2, Zip3, Zip4/Spo22, Mer3, Msh4, and Msh5, termed the SIC or ZMM proteins) that are also required for homologous chromosome synapsis. We report here genetic and molecular assays showing that sgs1 single mutants display relatively modest increases in CO recombination (less than 1.6-fold relative to wild-type). In contrast, a much greater CO increase is seen when an sgs1 mutation is introduced into the CO- and synapsis-deficient zip1, zip2, zip3, mer3, or msh4 mutants (2- to 8-fold increase). Furthermore, close juxtaposition of the axes of homologous chromosomes is restored. CO restoration in the mutants is not accompanied by significant changes in noncrossover (NCO) recombinant frequencies. These findings show that Sgs1 has potent meiotic anti-CO activity, which is normally antagonized by SIC/ZMM proteins. Our data reinforce previous proposals for an early separation of meiotic processes that form CO and NCO recombinants.


Transplantation of Allogeneic Pericytes Improves Myocardial Vascularization and Reduces Interstitial Fibrosis in a Swine Model of Reperfused Acute Myocardial Infarction.
Transplantation of adventitial pericytes (APCs) promotes cardiac repair in murine models of myocardial infarction. The aim of present study was to confirm the benefit of APC therapy in a large animal model. We performed a blind, randomized, placebo-controlled APC therapy trial in a swine model of reperfused myocardial infarction. A first study used human APCs (hAPCs) from patients undergoing coronary artery bypass graft surgery. A second study used allogeneic swine APCs (sAPCs). Primary end points were (1) ejection fraction as assessed by cardiac magnetic resonance imaging and (2) myocardial vascularization and fibrosis as determined by immunohistochemistry. Transplantation of hAPCs reduced fibrosis but failed to improve the other efficacy end points. Incompatibility of the xenogeneic model was suggested by the occurrence of a cytotoxic response following in vitro challenge of hAPCs with swine spleen lymphocytes and the failure to retrieve hAPCs in transplanted hearts. We next considered sAPCs as an alternative. Flow cytometry, immunocytochemistry, and functional/cytotoxic assays indicate that sAPCs are a surrogate of hAPCs. Transplantation of allogeneic sAPCs benefited capillary density and fibrosis but did not improve cardiac magnetic resonance imaging indices of contractility. Transplanted cells were detected in the border zone. Immunologic barriers limit the applicability of a xenogeneic swine model to assess hAPC efficacy. On the other hand, we newly show that transplantation of allogeneic sAPCs is feasible, safe, and immunologically acceptable. The approach induces proangiogenic and antifibrotic benefits, though these effects were not enough to result in functional improvements.


A novel feasible technique for rapid removal of broken proximal femoral nails: A case series.
Proximal femoral nail (PFN) fixation is a technique widely used to treat various femoral fractures, but femoral re-injury may occur, leading to PFN breakage. Broken nail removal, particularly removal of the distal broken nails, is usually challenging. Herein, we describe a feasible approach to successfully remove broken femoral intramedullary nails in three sites (proximal, middle, and distal section) of the PFN. Three patients required surgery for PFN breakage. We performed a novel technique using minimal exposure and a cerclage wire to remove the PFN fragment, which a distal knot on the wire was applied to hold the PFN fragment, and the removal trajectory was completed through the minimal exposure, a distal femoral bone window, and the marrow cavity. We successfully operated these three patients and removed the PFN fragments rapidly and effectively. In conclusion, this novel technique is rapid, feasible, and cost-effective, and can also be promising in removing intramedullary nail breakages in other long bones.


Pancreas beta-cells morphology, liver antioxidant enzymes and liver oxidative parameters in alloxan-resistant and alloxan-susceptible Wistar rats: a viable model system for the study of concepts into reactive oxygen species.
The aim of this study was to investigate biochemical and antioxidant parameters in alloxan-resistant (ALR) and alloxan-susceptible (ALS) rats. Diabetes was induced in 60-day-old male Wistar rats by a single intraperitonial injection of alloxan (AL, 150 mg/kg). Ten days after induction, a group of rats showed a significant decrease in glycemia. This group was named alloxan-resistant group. Susceptible rats showed a remarkable increase in the plasma lipid content, blood glucose and HbA1. Glycogen content in the liver decreased significantly in the ALS group (2.08 +/- 0.41 mg%) compared with ALR group (4.22 +/- 0.18). Aspartate aminotransferase and alanine aminotransferase activities were quantified in the plasma. Interestingly, ALR rats showed a decrease in both activities (42.1 +/- 6.11 and 21.7 +/- 5.54 U/mL) when compared with ALS rats (59.1 +/- 6.55 and 58.1 +/- 7.28 U/mL). The TBARS index was significantly increased in the ALS liver (0.38 +/- 0.08 nm/mg protein) when compared with the ALR liver (0.18 +/- 0.04). Superoxide dismutase and catalase activities in the ALR (230 +/- 13 and 131 +/- 15 U/mg protein) liver showed a marked increase when compared with the ALS liver (148 +/- 13 and 68 +/- 5 U/mg protein). The immunohistochemical and hematoxilin-eosin analysis also revealed that pancreatic islets of ALR rats display a different morphology amongst the groups. These results suggest an increased regenerative or recovery process in the ALR rat pancreatic islets and an increased hepatic antioxidant defenses in these group of alloxan-resistant rats.


Mixed Wastewater Coupled with CO2 for Microalgae Culturing and Nutrient Removal.
Biomass, nutrient removal capacity, lipid productivity and morphological changes of Chlorella sorokiniana and Desmodesmus communis were investigated in mixed wastewaters with different CO2 concentrations. Under optimal condition, which was 1:3 ratio of swine wastewater to second treated municipal wastewater with 5% CO2, the maximum biomass concentrations were 1.22 g L-1 and 0.84 g L-1 for C. sorokiniana and D. communis, respectively. Almost all of the ammonia and phosphorus were removed, the removal rates of total nitrogen were 88.05% for C. sorokiniana and 83.18% for D. communis. Lipid content reached 17.04% for C. sorokiniana and 20.37% for D. communis after 10 days culture. CO2 aeration increased intracellular particle numbers of both microalgae and made D. communis tend to be solitary. The research suggested the aeration of CO2 improve the tolerance of microalgae to high concentration of NH4-N, and nutrient excess stress could induce lipid accumulation of microalgae.


Sestrin 2 attenuates sepsis-associated encephalopathy through the promotion of autophagy in hippocampal neurons.
Sepsis-associated encephalopathy (SAE) has typically been associated with a poor prognosis. Although sestrin 2 (SESN2) plays a crucial role in metabolic regulation and the stress response, its expression and functional roles in SAE are still unclear. In the present study, SAE was established in mice through caecal ligation and puncture (CLP). The adeno-associated virus 2 (AAV2)-mediated SESN2 expression (ie overexpression and knockdown) system was injected into the hippocampi of mice with SAE, and subsequently followed by electron microscopic analysis, the Morris water maze task and pathological examination. Our results demonstrated an increase of SESN2 in the hippocampal neurons of mice with SAE, 2-16 hours following CLP. AAV2-mediated ectopic expression of SESN2 attenuated brain damage and loss of learning and memory functions in mice with SAE, and these effects were associated with lower pro-inflammatory cytokines in the hippocampus. Mechanistically, SESN2 promoted unc-51-like kinase 1 (ULK1)-dependent autophagy in hippocampal neurons through the activation of the AMPK/mTOR signalling pathway. Finally, AMPK inhibition by SBI-0206965 blocked SESN2-mediated attenuation of SAE in mice. In conclusion, our findings demonstrated that SESN2 might be a novel pharmacological intervention strategy for SAE treatment through promotion of ULK1-dependent autophagy in hippocampal neurons.


Dysprosium-sensitized chemiluminescence system for the determination of enoxacin in pharmaceutical preparations and biological fluids with flow-injection sampling.
A novel trivalence dysprosium(Dy(3+))-sensitized chemiluminescence method was developed for the first time for the determination of enoxacin (ENX) using flow-injection sampling based on the chemiluminescence (CL) associated with the reaction of the Dy(3+)-cerium(Ce(IV))-S(2)O(3) (2-)-ENX system and the Dy(3+)-MnO(4) (-) S(2)O(3) (2-)-ENX system. The analytical conditions for CL emission were investigated and optimized. The relationship between the CL intensity of ENX and its concentration has good linearity, with a correlation coefficient of 0.9984-0.9994. The limit of detection (LOD, 3sigma) was 0.20 ng/mL for the Dy(3+)-ENX-S(2)O(3)(2-)-Ce(IV)-H(2)SO(4) system and 0.22 ng/mL for the Dy(3+)-ENX-S(2)O(3)(2-)-MnO(4) (-)-HNO(3) system. The relative standard deviation (RSD, n = 11) was 1.8% for 11 determinations of 60 ng/mL ENX. The proposed method was applied to the analysis of ENX in injections, serum and urine samples with a recovery of 98%-105%. A possible mechanism for this sensitized CL reaction is discussed by comparing the CL spectra with the fluorescence emission spectra. The proposed method represents a wide linear range, high sensitivity and accuracy, and can be used for the routine determination of ENX in pharmaceutical preparations and biological fluids.


Bcl-2 expression in acute myeloblastic leukaemia: relationship with autonomous growth and CD34 antigen expression.
The bcl-2 gene encodes a mitochondrial protein that inhibits the onset of apoptosis induced by growth factor withdrawal or cytotoxic agents. Using quantitative flow cytometry and expressing bcl-2 levels as the number of molecules of equivalent soluble fluorochrome (MESF) per cell, we have shown that bcl-2 protein expression in the blast cells from patients with acute myeloblastic leukaemia (AML) is heterogeneous, but not related to FAB type. The blast cells from AML patients with the capacity to grow and survive autonomously in vitro were found to have higher bcl-2 MESF values than those that were dependent upon exogenous growth factors. We have previously reported that the blast cells from 70% of AML patients exhibit autonomous growth and autocrine growth factor production in vitro and that this has been shown to be an important indicator of poor prognosis in AML. High bcl-2 expression has also been associated with a low complete remission rate and poor survival in AML. In the patients whose blast cells exhibited autonomous growth, neutralisation of endogenous GM-CSF resulted in down-regulation of bcl-2 protein, whereas in blast cells from patients whose cells proliferated only in the presence of added growth factors, incorporation of recombinant human (rh) GM-CSF in the culture media resulted in up-regulation of bcl-2. Because CD34 positivity has been reported as another indicator of poor prognosis in AML, we compared bcl-2 expression in cases of CD34 positive AML, CD34 negative AML and CD34 positive normal bone marrow cells. Bcl-2 was found to be strongly expressed on the CD34+ normal bone marrow cells. The blast cells from CD34+ AML patients expressed significantly higher bcl-2 levels than CD34- AML patients. In five cases of CD34+ AML, the bcl-2 levels were determined on purified CD34+ and CD34- blast cell populations. The CD34+ blast cells were found to express significantly higher bcl-2 levels compared with the CD34-blast cells. Our data would suggest that quantification of bcl-2 in AML blast cell may be useful as a prognostic indicator in AML.


The Reverdin Graft: The Forgotten Skin Transplant.
This article describes a series of four patients for whom a Reverdin graft was performed. The Reverdin graft, also known as a pinch graft, is a method to promote epithelialization for superficial wounds. The intervention is minimally invasive with a short learning curve. The procedure and its advantages and disadvantages are discussed in this case series. This pinch graft is a widely accepted, minimally invasive intervention to accelerate the epithelialization of wounds.


Functional characterization of a cell wall invertase inhibitor StInvInh1 revealed its involvement in potato microtuber size in vitro.
Cell wall invertase (CWI) is as an essential coordinator in carbohydrate partitioning and sink strength determination, thereby playing key roles in plant development. Emerging evidence revealed that the subtle regulation of CWI activity considerably depends on the post-translational mechanism by their inhibitors (INHs). In our previous research, two putative INHs (StInvInh1 and StInvInh3) were expected as targets of CWI in potato (Solanum tubersum), a model species of tuberous plants. Here, transcript analysis revealed that StInvInh1 showed an overall higher expression than StInhInh3 in all tested organs. Then, StInvInh1 was further selected to study. In accordance with this, the activity of StInvInh1 promoter increased with the development of leaves in plantlets but decreased with the development of microtubers in vitro and mainly appeared in vascular bundle. The recombinant protein StInvInh1 displayed inhibitory activities on the extracted CWI in vitro and StInvInh1 interacted with a CWI StcwINV2 in vivo by bimolecular fluorescence complementation. Furthermore, silencing StInvInh1 in potato dramatically increased the CWI activity without changing activities of vacuolar and cytoplasmic invertase, indicating that StInvInh1 functions as a typical INH of CWI. Releasing CWI activity in StInvInh1 RNA interference transgenic potato led to improvements in potato microtuber size in coordination with higher accumulations of dry matter in vitro. Taken together, these findings demonstrate that StInvInh1 encodes an INH of CWI and regulates the microtuber development process through fine-tuning apoplastic sucrose metabolism, which may provide new insights into tuber development.


Solid-phase synthesis and CD spectroscopic investigations of novel beta-peptides from L-aspartic acid and beta-amino-L-alanine.
[structure: see text] A solid-phase synthesis method for the preparation of novel beta3- and beta2-peptides derived from l-aspartic acid and beta-amino-l-alanine, respectively, is described. The methodology allows independent buildup of the beta-peptide backbone and the introduction of sequential side chain substitutions. Representative peptides from the two classes, an amino-substituted beta3-hexapeptide and an acyl-substituted beta2-hexapeptide, have been prepared, and their solution conformation is studied by circular dichroism (CD) spectroscopy.


Atypical arterial supply to the spleen by polar branches of splenic artery and accessory splenic artery - a case report.
Vascular anomalies of the spleen are usually asymptomatic. However, variant anatomy of splenic artery becomes clinically important, when the patients undergo diagnostic angiography for gastrointestinal bleeding or during transcatheter therapy. We report here a concurrent variant arterial pattern of the spleen. The splenic artery was unusually elongated and excessively tortuous. Prior to its normal termination into segmental arteries, it gave superior and inferior polar arteries which entered the spleen distal to corresponding ends of the splenic hilum. In addition to this, the spleen also received an additional blood supply from an accessory splenic artery arising from left gastro-epiploic artery. The accessory splenic artery entered the substance of the spleen through its lateral end. Presence of such kind of concurrent variant arterial pattern of spleen makes the surgeons obligatory to have prior knowledge to prevent bleeding during any surgical or radiological interventional procedures targeting the spleen, pancreas etc.


Enzyme therapy for lysosomal storage disease: principles, practice, and prospects.
Over the past three decades, enzyme therapy for lysosomal storage diseases has moved from an academic pursuit to direct delivery of effective clinical care for affected patients and families. This success is based on understanding the complexities of lysosomal biogenesis, lysosomal hydrolase sorting and hydrolytic requirements, and the target sites of pathology of these diseases. This article reviews these concepts and their application to the treatment of affected patients with Gaucher disease, Fabry disease, and mucopolysaccharidosis I. The principles, progress, and practice in these diseases provide prototypes for expansion of enzyme therapy to a growing set of these diseases.


Posterior ankle impingement syndrome.
Posterior ankle impingement syndrome is a clinical disorder characterized by posterior ankle pain that occurs in forced plantar flexion. The pain may be acute as a result of trauma or chronic from repetitive stress. Pathology of the os trigonum-talar process is the most common cause of this syndrome, but it also may result from flexor hallucis longus tenosynovitis, ankle osteochondritis, subtalar joint disease, and fracture. Patients usually report chronic or recurrent posterior ankle pain caused or exacerbated by forced plantar flexion or push-off maneuvers, such as may occur during dancing, kicking, or downhill running. Diagnosis of posterior ankle impingement syndrome is based primarily on clinical history and physical examination. Radiography, scintigraphy, computed tomography, and magnetic resonance imaging depict associated bone and soft-tissue abnormalities. Symptoms typically improve with nonsurgical management, but surgery may be required in refractory cases.


Hysterectomy, oophorectomy, and cognitive function in older women.
It has been proposed that estrogen deficiency is a cause of memory loss in postmenopausal women. Bilateral oophorectomy leads to an abrupt decrease in gonadal hormone levels including testosterone. Hysterectomy with ovarian conservation may also lead to a decline in ovarian hormones. This study examines the effects of hysterectomy and oophorectomy on cognitive function in a large sample of older, postmenopausal women with and without current estrogen replacement therapy. Cross-sectional. Community-based sample in Southern California. Subjects were 885 postmenopausal women aged 60 to 89 who were initially enrolled in the Rancho Bernardo Study between 1972 and 1974 and had a follow-up clinic visit in 1988-91. All were Caucasian, middle to upper-middle class, relatively well educated, and ambulatory. Participants completed 12 standardized tests of cognitive function in 1988-91. Hysterectomy and oophorectomy status were assessed and use of estrogen therapy was also ascertained and validated. Of the 885 women, whose mean age +/- standard deviation was 74.3 +/- 7.5, 225 reported having had a hysterectomy with conservation of one or both ovaries and 190 reported having a hysterectomy with bilateral oophorectomy. Current estrogen use was reported by 35.4%. Among women not using estrogen, there were no significant differences on mean cognitive function scores by hysterectomy and oophorectomy status. Among current estrogen users, after adjustment for age, education, age at menopause, and past estrogen use, those with a hysterectomy and bilateral oophorectomy performed significantly less well on serial sevens and Trails B. These differences, although statistically significant, were small and unlikely to be of clinical significance. No differences in cognitive function were observed between women who had a hysterectomy with ovarian conservation and intact women. The present study suggests that, overall, there are no long-term effects of hysterectomy and bilateral oophorectomy on cognitive function and does not support the thesis that estrogen deficiency is associated with poor cognitive function in postmenopausal women.


Application of CEC procedures for the analysis of synthetic peptides: characterization of linear immunogenic peptides that mimic a HIV-1 gp120 epitope.
In this study, we describe the application of a new analytical procedure based on capillary electrochromatographic(CEC) techniques for the characterization of different basic and acidic peptides using isocratic eluent conditions containing acetonitrile and ammonium acetate buffers of different molarities between pH 3.8 and 5.2. In particular,10 immunogenic peptide analogs with isoelectric points ranging from 3.7 to 10.1 were investigated; nine of these peptides, 1-9, were truncated analogs of the parent peptide, 10, which is a peptidomimetic related to a HIV-1 gp120 epitope. Several of these peptides have the propensity to form alpha-helical secondary structures in solution. Electrochromatographic separations of these peptides were achieved with packed fused silica capillaries(25 cm packed length, 100 microm i.d.) containing 3 microm n-octadecylsilica particles. The influence of temperature on the CEC elution behavior of these peptides, as well as the impact of changes in the eluent composition, e.g. pH, buffer concentration and acetonitrile content, were examined. The results confirm that improvements in the resolution and analysis of synthetic peptides by CEC procedures result from the increase inelectroosmotic flow (EOF) as the temperature is increased. These findings emphasize the dominant influence of the temperature-dependent viscosity parameter, eta, on the EOF and thus on peptide resolution in CEC. Moreover, these investigations have shown that eluent properties can be specifically chosen to favor either electrophoretic mobility or chromatographic retention, with the overall CEC selectivity peptides of different sequence or composition reflecting the summated contributions from both separation mechanisms. Over the pH range 4.0-5.0, and using eluents with ionic strengths ranging from 6.2 to 15 mM ammonium acetate but containing a fixed volume fraction, psi, of acetonitrile above psi = 0.40, the CEC retention behavior of peptides 1-10 correlated with a linear relationship linking the retention coefficient, kappta(cec), and the differential frictional size-to-mass ratio parameter, Xi(fric), of these peptides. However, using eluents with a low acetonitrile content and low pH values, linear correlations were also observed between the incremental retention coefficient, Delta(Kappa)cec, and the product term [-0.66(Delta(Sigma[Xn]) log(Mi/Mj)], which links the difference in intrinsic hydrophobicities and molecular masses of two peptides, Pi and Pj. This study thus demonstrates the power of CEC procedures in the analysis of synthetic bioactive peptides and provides a general experimental framework to evaluate,using CEC procedures, the influence of the key molecular attributes of peptides on their structure-retention dependencies.Finally, these studies provide additional, practical insights into the use of CEC procedures for the analysis, resolution and biophysical characterization of closely related peptide analogs derived from solid-state peptide synthesis under conditions of different eluent composition or temperature.


Ultra-fast speech comprehension in blind subjects engages primary visual cortex, fusiform gyrus, and pulvinar - a functional magnetic resonance imaging (fMRI) study.
Individuals suffering from vision loss of a peripheral origin may learn to understand spoken language at a rate of up to about 22 syllables (syl) per second - exceeding by far the maximum performance level of normal-sighted listeners (ca. 8 syl/s). To further elucidate the brain mechanisms underlying this extraordinary skill, functional magnetic resonance imaging (fMRI) was performed in blind subjects of varying ultra-fast speech comprehension capabilities and sighted individuals while listening to sentence utterances of a moderately fast (8 syl/s) or ultra-fast (16 syl/s) syllabic rate. Besides left inferior frontal gyrus (IFG), bilateral posterior superior temporal sulcus (pSTS) and left supplementary motor area (SMA), blind people highly proficient in ultra-fast speech perception showed significant hemodynamic activation of right-hemispheric primary visual cortex (V1), contralateral fusiform gyrus (FG), and bilateral pulvinar (Pv). Presumably, FG supports the left-hemispheric perisylvian "language network", i.e., IFG and superior temporal lobe, during the (segmental) sequencing of verbal utterances whereas the collaboration of bilateral pulvinar, right auditory cortex, and ipsilateral V1 implements a signal-driven timing mechanism related to syllabic (suprasegmental) modulation of the speech signal. These data structures, conveyed via left SMA to the perisylvian "language zones", might facilitate - under time-critical conditions - the consolidation of linguistic information at the level of verbal working memory.


Benign mimickers of malignant breast lesions.
Breast pathology is filled with pitfalls, including underdiagnosis of bland-appearing lesions, both invasive and non-invasive, misdiagnosis of malignant lesions as belonging to the wrong subgroup, for example, calling LCIS as DCIS or missing the metaplastic component of an invasive lesion, and overdiagnosis of benign lesions as malignancy. While each is a sin of varying severity, the overdiagnosis of benign lesions can be the most scarring, especially in this age where Angelina Jolie׳s prophylactic mastectomy is the headline news and patients are pushing for aggressive preventive treatment. In this review, we will consider some of the more common benign lesions and the malignant counterpart that they mimic, with the goal of identifying characteristic features that will lead to the correct diagnosis. Much of the discussions will center around the assessment of core biopsies, as smaller tissue volume is most often contributory to overcalling benign lesions.


Myocardial regeneration strategies using human embryonic stem cell-derived cardiomyocytes.
Regenerative medicine is a new biomedicine discipline that takes advantage of the recent advancements in the fields of stem cell biology, molecular biology, and tissue engineering to derive tissue substitutes, in an attempt to replace or modify the function of diseased organs. The heart represents an attractive candidate for these emerging technologies since adult cardiac tissue has limited regenerative capacity. Consequentially, myocardial cell replacement therapy has emerged as a novel therapeutic paradigm for restoration of the myocardial electromechanical function. This innovative strategy has been significantly hampered, however, by the paucity of cell sources for human cardiomyocytes. The recent establishment of the human embryonic stem cell (hESC) lines may provide a possible solution for this cell-sourcing problem. These unique pluripotent cell lines can be propagated in the undifferentiated state in culture and coaxed to differentiate into cell derivatives of all three germ layers, including cardiomyocytes. This review will describe the hESC system, their differentiation into cardiomyocytes, and the structural and functional characterization of these cardiac lineage derivatives. The potential applications of this unique differentiating system in several research areas will be discussed with special emphasis on the steps required to fully harness their unique potential in the emerging field of cardiovascular regenerative medicine.


Histomorphometric analysis of iliac trabecular bone in otosclerosis.
With the aim of investigating whether patients with otosclerosis suffer from a generalized bone disorder, a histomorphometric analysis of trabecular bone from the iliac crest was performed. Iliac crest biopsies from 10 patients with otosclerosis and 20 normal controls were obtained after tetracycline double labelling and examined by histomorphometry. No significant differences were found in the estimates of density and remodeling of bone between patients and sex and age-matched normal controls. Qualitatively the bone specimens showed no signs of any generalized bone disorder.


The relationship between childhood adversity and dysphoric inner states among borderline patients followed prospectively for 10 years.
Childhood experiences of abuse and neglect were assessed in relation to dysphoric states among patients with borderline personality disorder (BPD) over a 10-year course of prospective follow-up. The Revised Childhood Experiences Questionnaire was administered at baseline to 290 patients meeting DIB-R and DSM-III-R criteria for BPD. The Dysphoric Affect Scale--a 50-item self-report measure of affective and cognitive states thought to be common among and specific to borderline patients--was administered at fives waves of prospective follow-up. Significant predictors of dysphoric states included emotional abuse, verbal abuse, physical abuse, sexual abuse, emotional withdrawal, inconsistent treatment, denial of patient's feelings, lack of a real relationship, placing patient in parental role, and failure to protect patient. This suggests that abusive and neglectful childhood experiences are significant risk factors for severe affective and cognitive difficulties reported by borderline patients and that sexual abuse is neither necessary nor sufficient for the development of these troubling inner states.


Resonant frequency characteristics of a SAW device attached to resonating micropillars.
Recently we reported experimental and simulation results on an increase in resonance frequency of a SAW resonator caused by mass loading of micropillars made of SU-8, attached normal to the surface of the resonator. We concluded that SAW resonator and the SU-8 micropillars in unison form a system of coupled resonators. We have now extended this work and performed a finite element method simulation to study the resonance frequency characteristics of the SAW-based coupled resonator. In this paper we report the effect of the resonance frequency of the micropillars on the resonance frequency of the system of coupled resonators, and observe the coupling of micropillar resonance and the propagating SAW as described in the well known Dybwad system of coupled resonators.


Influence of unilateral and bilateral knee extension fatigue protocols on inter-limb asymmetries assessed through countermovement jumps.
This study explores the sensitivity of jump type (unilateral and bilateral) and output variable (mean force, propulsive impulse, and jump height) to detect the changes in inter-limb asymmetries induced by unilateral and bilateral fatigue protocols. Thirty-eight individuals performed two testing sessions that consisted of (I) nine "pre-fatigued" countermovement jumps (CMJs; three bilateral and six unilateral [three with each leg]), (II) fatigue protocol and (III) nine "post-fatigued" CMJs. The testing sessions only differed in the fatigue protocol (five sets to failure against the 15-repetition maximum load using either the unilateral or bilateral knee extension exercise). The magnitude of all CMJ-derived variables (mean force, impulse, and jump height) decreased following both unilateral (p ≤ 0.002) and bilateral fatigue protocols (p ≤ 0.018). However, only unilateral protocol accentuated inter-limb asymmetries, which was detected for all variables during the unilateral CMJ (from -4.33% to -2.04%; all p < 0.05) but not during the bilateral CMJ (from -0.64% to 0.54%; all p > 0.05). The changes in inter-limb asymmetries following the unilateral and bilateral fatigue protocols were not significantly correlated between the unilateral and bilateral CMJs (rs ≤ 0.172). The unilateral CMJ should be recommended for the testing purposes over the bilateral CMJ due to its greater sensitivity to detect the selective effects of fatigue.


An 11-color flow cytometric assay for identifying, phenotyping, and assessing endocytic ability of peripheral blood dendritic cell subsets in a single platform.
Human peripheral blood dendritic cells (PBDC) are a rare population comprised of several distinctive subsets. Analysis of these cells has been hindered by their low frequency. In this study, we report a novel direct ex vivo 11-color flow cytometric assay that combines subset identification with analysis of activation status and endocytic ability of three major PBDC subsets (CD1c(+)CD11c(+) "MDC1," CD141(+)CD11c(+) "MDC2," and CD303(+)CD11c(-) "PDC") within a single platform. This method eliminates the need for DC enrichment, isolation, or prolonged culture. Human peripheral blood mononuclear cells (PBMC) from healthy donors are incubated with FITC-dextran directly ex vivo, prior to cell surface staining with various markers. As expected, PBDC identified by this assay express low levels of CD40 and CD86 directly ex vivo, and significantly upregulate expression of these molecules upon stimulation with toll-like receptor ligands LPS and CpG oligonucleotides. In addition, PDC internalize FITC-labeled dextran poorly in comparison to MDC1 and MDC2 subsets. Specificity of FITC-dextran endocytosis is further verified by imaging flow cytometry. Furthermore, the combination of surface markers used in this assay reveals a previously unreported CD4(+)CD11c(+)CD303(-)CD1c(-)CD141(-) cell population. Taken together, this assay is a rapid and cost-effective method that avoids manipulation of PBDC while providing direct ex vivo high-dimensional flow cytometry data for PBDC studies.


Reemerging nutritional rickets: a historical perspective.
Recent case reports highlight the resurgence of rickets in certain groups of breastfed infants. Infants residing in the North, irrespective of skin color, and dark-skinned African American infants residing anywhere in the United States are most vulnerable to nutritional rickets if they are exclusively breastfed past age 6 months without vitamin D supplementation. At the turn of the 20th century, rickets was nearly universal among African American infants living in the North. The discovery of vitamin D, the initiation of public health campaigns to fortify infant foods with vitamin D, and the supplementation of vitamin D to breastfed infants were responsible for overcoming the rickets scourge. We review a classic nutritional study by Alfred F. Hess, one of the greatest clinical nutritional researchers of the early 20th century, in the context of the resurgence of rickets, especially among dark-skinned infants. The Columbus Hill district, a black community of New York, NY, served as the setting for the study. Sixty-five infants (aged 1-17 months) entered a 6-month open-label trial of daily cod liver oil therapy. Participants were assessed for signs of rickets at recruitment and at 2, 4, and 6 months. Cod liver oil prevented the development of rickets in 34 (92%) of 37 infants treated for 6 months and in 7 (58%) of 12 treated for 4 months. Of the 16 infants who did not take the prescribed treatment, rickets progressed unremittingly in 15. Hess translated his success into a public health campaign leading to the development of the first rickets clinic in 1917. This was the first step in the conquest of the rickets epidemic of the early 20th century.


Salivary proteomics in monitoring the therapeutic response of canine oral melanoma.
Saliva biomarkers are suitable for monitoring the therapeutic response of canine oral melanoma (COM), because saliva directly contacts the tumor, and saliva collection is non-invasive, convenient and cost effective. The present study aimed to investigate novel biomarkers from the salivary proteome of COM treated with surgery and a chemotherapy drug, carboplatin, 1-6 times, using a liquid chromatography-tandem mass spectrometry approach. The expression of a potential salivary biomarker, ubiquitin D (UBD), was observed and verified by western blot analysis. A significantly increased ratio of free UBD (fUBD) to conjugated UBD (cUBD) was shown in the pre-surgery stage (PreS) in OM dogs with short-term survival (STS) (less than 12 months after surgery) compared with that with long-term survival (more than 12 months after surgery). In dogs with STS, the ratio was also shown to be augmented in PreS compared with that after surgery, followed by treatment with carboplatin twice, 4 and 5 times [After treatment (AT)2, AT4 and AT5]. In addition, the expression of fUBD was enhanced in PreS compared with that of AT2 in the STS group. In conclusion, this study revealed that a ratio of fUBD to cUBD in PreS was plausibly shown to be a potential prognostic biomarker for survival in dogs with OM.