[Myotoxicity and rhabdomyolisis due to statins].
Rhabdomyolysis is a rare but potentially fatal complication associated with the use of cholesterol synthesis inhibitors (statins). The complication can develop in susceptible patients and with the concomitant use of medications that impede the biodegradation of statins, for example, biotransformation via the cytochrome P450 system. This may result in the plasma and tissue concentrations of statins, and their active metabolites, increasing to levels that are toxic for striated muscle. Myopathy is present when plasma activity levels of creatinine kinase are raised to in excess of 10 times the upper limit of the normal value. Muscular complaints which may be indicative of myotoxicity and subsequent myopathy are present in 1-7% of statin users. Albeit to varying degrees, all statins can induce myotoxicity especially at high dosages. Rhabdomyolysis was clearly more prevalent under cerivastatin users than the users of other statins and was therefore recently withdrawn from the market. Statins should be withdrawn immediately if myopathy is suspected. Prompt withdrawal may prevent rhabdomyolysis.


On-the-Fly Computation of Frontal Orbitals in Density Matrix Expansions.
We propose a method for computation of frontal (homo and lumo) orbitals in recursive polynomial expansion algorithms for the density matrix. Such algorithms give a computational cost that increases only linearly with system size for sufficiently sparse systems, but a drawback compared to the traditional diagonalization approach is that molecular orbitals are not readily available. Our method is based on the idea to use the polynomial of the density matrix expansion as an eigenvalue filter giving large separation between eigenvalues around homo and lumo [ Rubensson et al. J. Chem. Phys. 2008 , 128 , 176101 ]. This filter is combined with a shift-and-square (folded spectrum) method to move the desired eigenvalue to the end of the spectrum. In this work we propose a transparent way to select recursive expansion iteration and shift for the eigenvector computation that results in a sharp eigenvalue filter. The filter is obtained as a byproduct of the density matrix expansion, and there is no significant additional cost associated either with its construction or with its application. This gives a clear-cut and efficient eigenvalue solver that can be used to compute homo and lumo orbitals with sufficient accuracy in a small fraction of the total recursive expansion time. Our algorithms make use of recent homo and lumo eigenvalue estimates that can be obtained at negligible cost [ Rubensson et al. SIAM J. Sci. Comput . 2014 , 36 , B147 ]. We illustrate our method by performing self-consistent field calculations for large scale systems.


Combined Hepatic and Pulmonary Metastasectomies From Colorectal Carcinoma. Data From the Prospective Spanish Registry 2008-2010.
Resection of both liver and lung metastases from colorectal carcinoma (CRC) is a standard of care in selected patients with oligometastatic disease. We present here the analysis of the subgroup of patients undergoing combined surgery from the Spanish Group of Surgery of Pulmonary Metastases (PM) from Colorectal Carcinoma (GECMP-CCR-SEPAR). We analyze characteristics, survival and prognostic factors of patients undergoing combined resection from March-2008 to February-2010 and followed-up during at least 3 years, from the prospective multicenter Spanish Registry. A total of 138 patients from a whole series of 543 cases from 32 thoracic surgery units underwent both procedures. Seventy-seven (43.8%) resected liver metastases were synchronic with colorectal tumor. Median disease specific survival (DSS) from first pulmonary metastasectomy was 48.9 months, being three and 5-year DSS 65.1% and 41.7%, respectively. From CRC-surgery median DSS was 97.2 months, with 3 and 5-year DSS rates of 96.7% and 77%, respectively. Five-year DSS from pulmonary metastasectomy was 41.7% for patients with combined resection and 52.4% for those without hepatic involvement (P=.04). Differences disappeared when considering DSS from colorectal surgery. Carcinoembrionary antigen (CEA) before lung surgery over 10mg/dl and bilateral PM were independent prognostic factors for survival (hazard ratio 2.4 and 2.5, respectively). Patients with resection of PM of CRC with history of resected hepatic metastases presented significantly lower disease specific survival rates than those undergoing pulmonary metastasectomy alone. CEA before lung surgery and bilateral PM associated worse prognosis.


The mechanism of diflufenican resistance and its inheritance in oriental mustard (Sisymbrium orientale L.) from Australia.
An oriental mustard population (P3) collected near Quambatook, Victoria was identified as being resistant to diflufenican by screening with the field rate (200 g a.i. ha-1 ) of the herbicide. The mechanism(s) of diflufenican resistance and its inheritance in this population were therefore investigated. Dose-response experiments confirmed that population P3 was 140-fold more resistant to diflufenican than susceptible populations, as determined by the comparison of 50% lethal (LD50 ) values. The phytoene desaturase (PDS) gene from five individuals each of the S1 [susceptible (S)] and P3 [resistant (R)] populations was sequenced, and a substitution of valine for leucine at position 526 (Leu-526-Val) was detected in all five individuals of P3, but not in the S1 population. Inheritance studies showed that diflufenican resistance is encoded in the nuclear genome and is dominant, as the response to diflufenican at 200 g a.i. ha-1 of F1 families was equivalent to that of the resistant biotype. The segregation of F2 phenotypes fitted a 3:1 inheritance model. Segregation of 42 F2 individuals by genotype sequencing fitted a 1:2:1 (ss:Rs:RR) ratio. Resistance to diflufenican in oriental mustard is conferred by the Leu-526-Val mutation in the PDS gene. Inheritance of resistance is managed by a single gene with high levels of dominance. © 2018 Society of Chemical Industry.


[Genic alterations in oral and head and neck squamous cell carcinomas: analysis of international literature].
The development of oral and head and neck squamous cell carcinomas occurs in relation with multiple events including mainly: loss of cycle cell control, evasion from apoptosis, telomerase reactivation. Complex interactions between a set of molecules, cell cycle proteins, tumour suppressor genes, oncogenes and the telomerase, occur in the multiple step process of carcinogenesis. The 2 main ways of control of the cell cycle rely on 2 tumour suppressor genes: the P53 gene and the retinoblastoma gene or RB gene. One of the regulation pathways or the 2 regulation pathways are disabled during the development of oral and head and neck squamous cell carcinomas. Most of the time, the inactivation of the P53 pathway results from a loss of function of the p53 protein, secondary to mutation and/or deletion of the P53 gene; It may also result of the amplification of the MDM2 gene and of the inactivation of the arf protein. The RB pathway leads to cell proliferation by loss of the p16 protein, by amplification of the cyclin D1 gene and less frequently by mutation of the RB gene or loss of the retinoblastoma protein. In India and South-East Asia, the activation of RAS and MYC oncogenes appears to be related with the presence of specific carcinogens in snuff and tobacco. By blocking apoptosis, the Bcl2 protein seems to increase the resistance of tumours to radiotherapy and chemotherapy.


Facile assembly of bifunctional, magnetically retrievable mesoporous silica for enantioselective cascade reactions.
An integrated immobilization to encapsulate the Pd/C-coated magnetic nanoparticles within the chiral Ru/diamine-functionalized silica shell for the construction of a bifunctional magnetic catalyst is developed. This catalyst realizes a synergistic Suzuki cross-coupling/asymmetric transfer hydrogenation and a successive reduction/asymmetric transfer hydrogenation for the preparation of chiral aromatic alcohols.


In vitro alterations in cytoskeletal tensional homeostasis control gene expression in tendon cells.
An in vitro collagen gel system was used to determine the effect of alterations in cytoskeletal tensional homeostasis on gene expression in tendon cells. Collagen gel matrices, seeded with rat tail tendon cells, underwent cytochalasin D and gel contraction treatments designed to alter the internal cytoskeletal homeostasis of the cells. Gels were examined for cytoskeletal organization using a rhodamine phalloidin stain for actin. The effect of altered cytoskeletal organization on mRNA expression of a catabolic (interstitial collagenase) and anabolic (alpha1(I) collagen) gene was examined using northern blot analysis. Tendon cells in adhered gels demonstrated a highly organized cytoskeleton and showed evidence of alpha1(I) collagen mRNA expression but no evidence of collagenase mRNA expression. Treatment of the attached gel with cytochalasin D disrupted the cytoskeletal organization and resulted in the up-regulation of collagenase mRNA and the inhibition of alpha1(I) collagen mRNA expression. Release of the gels resulted in a cell mediated gel contraction, an immediate loss of cytoskeletal organization, and an mRNA expression pattern similar to that seen with cytochalasin D treatment. Isometric contraction of the gel on itself or around a 3-point traction device resulted in an mRNA expression pattern similar to the adhered gel. Gene expression in the contracted gels could be reversed through chemical cytoskeletal disruption or removal of the traction device which permitted further gel contraction. The results of the study suggest that tendon cells can establish an internal cytoskeletal tension through interactions with their local extracellular environment. Alterations in this tension appear to control the expression of both catabolic and anabolic genes in a reciprocal manner.


Pulse-dose radiofrequency can reduce chronic pain in trapezio-metacarpal osteoarthritis: A mini-invasive therapeutic approach.
To assess the efficiency of pulse-dose radiofrequency (PDRF) in the management of chronic pain in patients with trapezio-metacarpal osteoarthritis (OA). Seventy-five patients with trapezio-metacarpal OA were treated with the PDRF between October 2011 and September 2013. A 22-gauge cannula with a 5-cm length was introduced with percutaneous access in the lateral region of the affected trapezio-metacarpal joint. The PDRF procedure was performed with 1200 pulses at 45 V and 20 msec duration, followed by a 480 msec silent phase. The follow-up visits were performed at 1 month, 3 and 6 months after the PDRF procedure. All patients underwent a second treatment between 7 and 9 months after the first PDRF procedure with a new follow-up scheme at 1, 3, 6 and 9 months. Mean visual analogue scale (VAS) scores before the procedures was 8.5 ± 1.1. A great reduction in pain intensity was reported at 3 months after the PDRF procedure (mean VAS scores 3.1 ± 0.9, P < 0.05). Pain intensity began to increase back to pre-procedural values after 4 months (mean VAS scores at 6 months was 7.9 ± 1.3, P < 0.05). After the second PDRF treatment the mean VAS scores decreased to 3.3 ± 0.8 at 3 months, but increased to 8.1 ± 1.6 at 9 months. (P < 0.05). No complications after the procedures were observed. The PDRF may be a safe, repeatable and effective short-term pain management technique in patients with trapezio-metacarpal OA. Larger, randomized controlled studies are indicated to better clarify the efficacy and utility of the PDRF.


Type I interferons and the quality of life of multiple sclerosis patients. Results from a clinical trial on interferon alfa-2a.
The objective of the study was to examine whether the beneficial effect of treatment of interferon alfa-2a on multiple sclerosis seen by magnetic resonance imaging is reflected in a corresponding improvement in the quality of life (QoL) and to address the impact of adverse events related to this treatment on the QoL. The study was a randomised double-blinded placebo-controlled treatment trial including 97 relapsing-remitting multiple sclerosis patients. Thirty-two patients received 4.5 MIU recombinant interferon alfa-2a, 32 patients received 9.0 MIU recombinant interferon alfa-2a and 33 patients received placebo treatment for 6 months. All patients were followed up 6 months after end of treatment. QoL was assessed according to the eight scales of the SF-36 Health Survey and measured at baseline, month 3, 6 and 12. The effect found on MRI was not reflected in a corresponding change in the QoL. We found a relationship between the presence of new enhancing lesions and reduced QoL among the placebo patients, whereas this was not found among the patients treated with interferon. The presence of the adverse events fatigue, myalgia, headache and weakness were significantly negatively correlated to several of the QoL dimensions. Conclusively, the treatment with interferon alfa-2a does not seem to improve the patients' QoL after 6 months of treatment, in spite of a marked effect measured by MRI. The treatment is followed by adverse events that negatively affected the QoL.


Echo-planar microscopy of porous rocks.
A modified version of the echo-planar imaging sequence incorporating multiple 180 degrees RF pulses (PEPI) has been implemented at 11.7 T and used to generate high resolution images of porous solids. By preceding the PEPI sequence with an inversion recovery or flow-encoding sequence, rapid T1, and velocity mapping is possible in the microscopic domain.


Multi-site study of the Childhood Autism Rating Scale (CARS) in five clinical groups of young children.
This study examined several questions pertaining to the Childhood Autism Rating Scale (CARS) in a sample of 274 preschool children (aged 2-6 years) clinically diagnosed as falling in one of five groups: Autistic Disorder, PDD-NOS, MR, Delayed, and Other. In addition to diagnosis and the CARS, all children were given standardized cognitive and adaptive behavior measures. Results indicated high concordance between the CARS and clinical diagnosis using DSM-IV (including excellent sensitivity and specificity). There was a moderate negative correlation of CARS scores and developmental level (both cognitive and adaptive), indicating significant shared variance. There were significant and sensible differences in mean CARS score for different diagnostic groups, including a substantial difference between the Autistic Disorder and PDD-NOS groups.


[Comparison of normal tension and high tension glaucoma patients (corrected) according to age and sex].
According to the level of intraocular pressure (IOP), open angle glaucoma is divided into high tension glaucoma (HTG) and normal tension glaucoma (NTG). To determine if there are differences in the distribution of patients by age and sex between NTG and HTG. Our prospective study included 30 patients with NTG and 30 with HTG. A complete eye examination was performed. The examination included measuring of intraocular pressure by Goldmann applanation tonometry, examination of optic disc head by indirect ophthalmoscopy with Volk 90 D lens and visual field examination using the Octopus program. The average damage of visual field in the group of patients with NTG was 8.14 +/- 4.43 dB, while in the group with HTG it was 7.40 +/- 2.84 dB (p > 0.05). The average age of the group of patients with NTG was 66 +/- 11.58 years, while among those with HTG the average age was 59.7 +/- 11.63 years (p < 0.01). Among the patients with NTG there were three times more women than men (chi2 = 9.124; p < 0.01), while in the group of patients with HTG there were more men than women, but without statistically significant difference between the tested groups (chi2 = 1.851; p > 0.05). Open angle glaucoma is a disease of elderly population. According to our results, risk factors for this disease can be age and sex. NTG is more frequently present than HTG among elderly population and females.


[Chinese material medica combined with cisplatin and lipiodol through transcatheter arterial embolization in the treatment of primary hepatoma].
Transcatheter arterial embolization (TAE) using hydroxycamptothecin, cantharidin and cisplatin which were mixed thoroughly with lipiodol, combined with large doses interferon and interleukin-2 as adoptive immunotherapy were carried out in the treatment of 48 patients with unresectable advanced stage primary hepatoma, evaluation of therapeutic effect showed that partial remission rate was 54.2%, significantly higher than that of embolization group using chemotherapeutic agents alone (cisplatin, adriamycin and mitomycin), the partial remission rate was 32.1% (P < 0.01). The side effects of camptothecin and cantharidin including hematuria, urodynia were also successfully eliminated.


[Complex treatment of elderly patients with pain reflectory manifestations of lumbosacral osteochondrosis and peripheral circulation disorder].
Involutional age makes difficult the treatment of pain syndromes of vertebra origin. Concomitant diseases, age stress syndrome, hormone disbalance constrain the management of the disease. Combination of laser stimulation, reflexotherapy, psychodiagnostics and psychocorrection enhances the efficiency of the treatment of pain syndromes in patients with lumbo-sacral radiculitis and peripheral circulation disorder.


[Diagnosis and treatment of severe hyperlipidemia].
Though severe hyperlipidaemia (total cholesterol level > or = 13 mmol/l in this study) is uncommon, it is important to make a precise diagnosis. We examined 57 patients with isolated severe hypercholesterolaemia. Of these, four were homozygotes for familial hypercholesterolaemia, 48 were heterozygotes for familial hypercholesterolacmia and one had sitosterolemia. The heterozygotes carried 15 different LDL receptor mutations, with no one mutation predominating. When the diagnosis is made, relatives should be given the opportunity to be tested. Combined severe hyperlipidaemia is usually due to a secondary cause, at our clinic, the most common cause is diabetes mellitus. The underlying disease should be treated first. However, many patients will require additional lipid-lowering drugs because the underlying disease may be associated with an increased risk of cardiovascular disease. With the exception of fish oil capsules, drugs that reduce serum triglyceride levels substantially are not registered in Norway at present.


Enzyme-assisted waste-to-reactant transformation to engineer renewable DNA circuits.
To date, implementation of renewable DNA circuits remains challenging due to issues including reactant depletion and waste accumulation. Herein we simultaneously addressed both issues through nicking enzyme-assisted waste-to-reactant transformation. As a proof-of-concept, a renewable entropy-driven catalytic DNA circuit was implemented, exhibiting a good renewability when replenishing fuel.


Photosynthetic adaptation to high temperatures: a field study in death valley, california.
The photosynthesis of Tidestromia oblongifolia (Amranthaceae) is remarkably well adapted to operate at the very high summer temperatures of the native habitat on the floor of Death Valley. The photosynthetic rate was very high and reached its daily maximum when the light intensity reached its noon maximum at the high leaf temperatures of 460 degrees to 50 degrees C which occurred at this time. At the intensity of noon sunlight the rate decreased markedly when the leaf temperature was experimentally reduced to below 44 degrees C. The optimum rate occurred at 47 degrees C. At this temperature the photosynthetic rate was essentially directly proportional to light intensity up to full sunlight.


Carbon monoxide exposure assessment among toll operators in Klang Valley, Kuala Lumpur, Malaysia.
A comparative cross-sectional study was conducted to determine tollbooth carbon monoxide (CO) levels and carboxyhaemoglobin (COHb) levels among the tollbooth operators and office workers in the Klang Valley, Kuala Lumpur. All tollbooths were equipped with well functioning air-conditioning. The total number of respondents was 180: 90 toll operators and 90 office workers aged between 19 and 52 years. The highest peak of CO level recorded was 61 ppm. The highest average peak CO level within a shift was 30 ppm. The CO level was higher during peak traffic at 6.00 - 8.00 a.m. There was no significant correlation between average peak CO level with vehicle load (r = -0.007, p = 0.474). The toll operators' median COHb level (1.0%, IQR = 0.8%) was significantly higher (p = 0.008) compared to office workers (0.7%, IQR = 0.8). There was a weak and significant correlation between COHb levels with average peak CO levels (r = 0.228, p = 0.031). In conclusion, tollbooth operators were chronically exposed to CO leading to higher COHb levels compared to office workers.


[Cholelithiasis in homozygous sickle cell anemia. Review of the literature].
Cholelithiasis in homozygous sickle cell disease is a common complication. Mean prevalence rate recorded is 27.4% with a greater frequency in the second part of life. It is a peculiar lithiasis, made of pigmentary stones either purs or duals, caused by hypersaturation of indirect bilirubin. Systematic detection of such a lithiasis by echography is part of monitoring homozygous drepanocytic.


The fiber orientation in the coronary arterial wall at physiological loading evaluated with a two-fiber constitutive model.
A patient-specific mechanical description of the coronary arterial wall is indispensable for individualized diagnosis and treatment of coronary artery disease. A way to determine the artery's mechanical properties is to fit the parameters of a constitutive model to patient-specific experimental data. Clinical data, however, essentially lack information about the stress-free geometry of an artery, which is necessary for constitutive modeling. In previous research, it has been shown that a way to circumvent this problem is to impose extra modeling constraints on the parameter estimation procedure. In this study, we propose a new modeling constraint concerning the in-situ fiber orientation (β (phys)). β (phys), which is a major contributor to the arterial stress-strain behavior, was determined for porcine and human coronary arteries using a mixed numerical-experimental method. The in-situ situation was mimicked using in-vitro experiments at a physiological axial pre-stretch, in which pressure-radius and pressure-axial force were measured. A single-layered, hyperelastic, thick-walled, two-fiber material model was accurately fitted to the experimental data, enabling the computation of stress, strain, and fiber orientation. β (phys) was found to be almost equal for all vessels measured (36.4 ± 0.3)°, which theoretically can be explained using netting analysis. In further research, this finding can be used as an extra modeling constraint in parameter estimation from clinical data.


Re-appraisal of malignant melanoma diagnosis in the Swedish cancer registry.
In many countries the registration of malignant melanoma of the skin shows an increased incidence; this may partially be due to an over-diagnosis. From the group of 3,268 adults and 21 children classified as suffering from malignant melanoma of the skin in the Swedish Cancer Registry between 1959-1968, a one-in-ten systematic sample survey was the object of a retrospective examination. 349 adults and 2 children had a uniform histopathological evaluation performed by one of the authors (B.L.). Slides were not available in 8 cases (2 per cent) and in the remaining 343, the diagnosis was regarded as incorrect in 13 (3.7 per cent). All these 13 cases, (2 children and 11 adults), showed no special predilection as to age, sex, location, or year of registration. In view of such a slight degree of error, the Swedish Cancer registry's melanoma group from 1959-1968 would seem suitable for further epidemiological studies.


Spreadability of two glass ionomer cements used in atraumatic restorative treatment (ART).
'Press finger' in atraumatic restorative treatment (ART) is used to spread a glass ionomer cement (GIC) to seal the restoration margin and adjacent pits and fissures. This study compared the spreadability of Fuji IX and Ketac-Molar to establish which was best suited for this purpose. Twenty equally spaced cavities (35 mm apart) were machine-cut in each of two Perspex slabs 200 x 100 x 20 mm. Each cavity was 3 mm deep with a 4 mm diameter. Two V-shaped grooves 1 mm deep were cut to traverse the cavities: one 0.5 mm wide, the other 0.25 mm wide. Equal amounts (0.25 ml) of GIC were dispensed, 20 cavities for Fuji IX and 20 for Ketac-Molar and condensed under light finger pressure using a 20 mm diameter cork thinly coated with petroleum jelly. This allowed the GIC to be condensed into the cavity and spread into the differently sized grooves. The samples were stored in deionised water for 24 hours whereafter the length to which the GIC had spread along each groove from the cavity edge was measured to the closest 0.01 mm. The data were analysed using ANOVA and the unpaired Student's t-test (P < 0.05). There was a statistically significant difference between the length of spread of the two GICs (t = 2.534; P = 0.013) which was confined to the 0.25 mm width groove (t = 2.83; P = 0.007) with Fuji IX spreading much further along the groove (10.25 +/- 1.17 mm) than Ketac-Molar (7.66 +/- 4.21 mm). Fuji IX appears to be the better sealant material when selecting for spreadability in ART.


Improved performance of a fully gutted adenovirus vector containing two full-length dystrophin cDNAs regulated by a strong promoter.
Dystrophin gene transfer using gutted or helper-dependent adenoviruses (HDAd), which have most of their genes deleted, is a promising approach to treat Duchenne muscular dystrophy. In an attempt to boost the amount of dystrophin produced after gene transfer, we have constructed a fully deleted HDAd (HDCBDys2x) containing two human dystrophin cDNAs controlled by the powerful hybrid cytomegalovirus enhancer/beta-actin promoter. We demonstrated high dystrophin expression after infection of muscle cultures with HDCBDys2x. Similarly, high (mean=583) and moderate (mean=124) numbers of muscle fibers were transduced in anterior tibialis muscle after intramuscular injection of HDCBDys2x in neonate and adult dystrophindeficient (mdx) mice 10 days postinjection. In fact, in the neonatally injected mdx mice, the transferred dystrophin was five times more abundant than in normal human muscle. However, the high early transduction level was transient in both animal groups, and we observed a humoral response against the human dystrophin. In contrast, we demonstrated sustained dystrophin expression in immunodeficient mouse muscles. Dystrophin expression of HDCBDys2x could be further increased in the presence of an E1/E3-deleted (first-generation) adenovirus, thus demonstrating that the latter vector synthesizes trans-acting enhancing factors. We have achieved abundant dystrophin expression with our new, improved HDAd. It is anticipated that high longterm transgene expression will be possible by employing weaker immunogenic transgenes.


Cytotoxic and antioxidant capacity of camel milk peptides: Effects of isolated peptide on superoxide dismutase and catalase gene expression.
Peptides from natural sources such as milk are shown to have a wide spectrum of biological activities. In this study, three peptides with antioxidant capacity were identified from camel milk protein hydrolysate. Pepsin and pancreatin were used for hydrolysis of milk proteins. Ultrafiltration and reverse-phase high-performance liquid chromatography were used for the concentration and purification of the hydrolysate, respectively. Sequences of the three peptides, which were determined by matrix-assisted laser desorption/ionization time-of-flight spectrophotometry, were LEEQQQTEDEQQDQL [molecular weight (MW): 1860.85 Da, LL-15], YLEELHRLNAGY (MW: 1477.63 Da, YY-11), and RGLHPVPQ (MW: 903.04 Da, RQ-8). The 3-(4,5-dimethylthia-zol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to evaluate the cytotoxicity of these chemically synthesized peptides against HepG2 cells. In vitro analysis showed antioxidant properties and radical scavenging activities of these peptides on 2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)+, O2-, and OH- free radicals. HepG2 cells were treated with YY-11 peptide for 48 hours, and the expression of superoxide dismutase and catalase genes was examined using real-time polymerase chain reaction. The results revealed a significant increase in the expression of superoxide dismutase and catalase genes in treated HepG2 cells.


Access to common laparoscopic general surgical procedures: do racial disparities exist?
Laparoscopy has become the standard of care for the majority of cases for inguinal hernia repair, cholecystectomy, appendectomy, and colectomy due to the shortened patient recovery time compared to open surgery. This study sought to determine if there exists racial disparity in access to a laparoscopic approach to these common surgeries. This was an IRB-approved retrospective study utilizing data from the American College of Surgeons National Surgical Quality Improvement Program (ACS-NSQIP). Individuals who underwent inguinal hernia repair, cholecystectomy, appendectomy, and colectomy in 2016 were identified. Information on self-reported race and ethnicity and other demographic and pre-operative clinical covariates were recorded. Propensity matching was conducted to evaluate the association between race and a laparoscopic approach to surgery. There were 44,522, 60,444, 50,523, and 58,012 cases of inguinal hernia repair, cholecystectomy, appendectomy, and colectomy identified, respectively. Of these patients, 8.38, 8.76, 6.69, and 9.02% self-identified as black, respectively. Confounding effects of variables other than race were balanced by propensity matching. After propensity matching, there were 7460, 10,574, 10,470, and 6758 cases of hernia repair, cholecystectomy, colectomy, and appendectomy, respectively. On univariate (Chi square) analysis with laparoscopic surgery as the primary outcome, black race was significantly associated with lower likelihood of undergoing a minimally-invasive surgical approach in all four surgical procedures under investigation (33.86% of white patients and 21.69% of black patients, p < 0.0001 for hernia repair; 97.98% of white patients and 94.29%, p < 0.0001 of black patients for cholecystectomy; 70.93% of white patients and 48.60% of black patients, p < 0.0001 for colectomy; and 98.85% of white patients and 92.81% of black patients, p < 0.0001 for appendectomy). There appears to be a significant racial disparity in the application of a laparoscopic approach to routine intra-abdominal surgery. This warrants further investigation into the barriers preventing access to laparoscopic general surgical procedures that certain populations face.


Chronic low back pain and disability in Brazilian jiu-jitsu athletes.
To identify the prevalence of chronic low back pain (CLBP) and functional disability in Brazilian jiu-jitsu athletes. Cross-sectional, observational. The study was conducted at jiu-jitsu training sites in the State of Rio de Janeiro. Presence of Chronic low back pain and Quebec Back Pain Disability Scale. The sample was composed of 72 athletes (mean age of 26.7), being 36 recreational and 36 professional. Chronic low back pain was present in 80.6% of athletes. Pain was present in 88.9% of professional and 72.2% of recreational athletes. In the professional jiu-jitsu group, the median of the Quebec Back Pain Disability Scale (QBPDS) was 10 (IQR = 16), and in the recreational group the QBPDS result was 6.0 (IQR = 12) (p = .001). Professional athletes had a marginally significant increased risk of developing CLBP [OR = 3.0; CI(95%) 0.8-10.9)]. The prevalence of low back pain in jiu-jitsu practice was high and professional athletes seem to have a high risk of developing CLBP.


Using age, triage score, and disposition data from emergency department electronic records to improve Influenza-like illness surveillance.
Markers of illness severity are increasingly captured in emergency department (ED) electronic systems, but their value for surveillance is not known. We assessed the value of age, triage score, and disposition data from ED electronic records for predicting influenza-related hospitalizations. From June 2006 to January 2011, weekly counts of pneumonia and influenza (P&I) hospitalizations from five Montreal hospitals were modeled using negative binomial regression. Over lead times of 0-5 weeks, we assessed the predictive ability of weekly counts of 1) total ED visits, 2) ED visits with influenza-like illness (ILI), and 3) ED visits with ILI stratified by age, triage score, or disposition. Models were adjusted for secular trends, seasonality, and autocorrelation. Model fit was assessed using Akaike information criterion, and predictive accuracy using the mean absolute scaled error (MASE). Predictive accuracy for P&I hospitalizations during non-pandemic years was improved when models included visits from patients ≥65 years old and visits resulting in admission/transfer/death (MASE of 0.64, 95% confidence interval (95% CI) 0.54-0.80) compared to overall ILI visits (0.89, 95% CI 0.69-1.10). During the H1N1 pandemic year, including visits from patients <18 years old, visits with high priority triage scores, or visits resulting in admission/transfer/death resulted in the best model fit. Age and disposition data improved model fit and moderately reduced the prediction error for P&I hospitalizations; triage score improved model fit only during the pandemic year. Incorporation of age and severity measures available in ED records can improve ILI surveillance algorithms.


Utilization of quercetin and quercetin glycosides from onion (Allium cepa L.) solid waste as an antioxidant, urease and xanthine oxidase inhibitors.
This study aimed to determine the flavonol glycosides from onion solid waste (OSW) using HPLC analysis, with antioxidant and enzyme inhibitory activities. We found considerable amount of quercetin-4'-O-monoglucoside (QMG: 254.85), quercetin-3,4'-O-diglucoside (QDG: 162.34), quercetin (Q: 60.44), and isorhamnetin-3-glucoside (IMG: 23.92) (mg/100g) dry weight (DW) of OSW. For OSW, the methanol and ethanol showed the strongest antioxidant activities, followed by ethyl acetate, chloroform, and n-hexane extracts. Among the flavonols, Q and QDG possessed higher antioxidant activities. OSW and flavonol glycosides displayed significant enzyme inhibitory activity, with IC50 values ranging from 12.5±0.11 to 32.5±0.28 for OSW, 8.2±0.07 to 16.8±0.02 for flavonol glycosides, and 4.2±0.05μg/mL for thiourea (positive control) towards urease; while 15.2±0.8 to 35.8±0.2 (μg/mL) for OSW, 10.5±0.06 to 20.8±0.05 (μg/mL) for flavonol glycosides, and 6.5±0.05μg/mL for allopurinol (positive control) towards xanthine oxidase, respectively. The OSW and flavonol glycosides may thus be considered as potential antioxidant and antigout agents.


[Reverse genetics platform construction of influenza pandemic virus strain].
Reverse genetics was used to construct the platform of flu pandemic strain A/California/07/2009 (H1N1). Eight genes fragements were amplified and ligated with bidirectional vector, recombinant plasmids were co transfected to the 293 T cells and rescued the virus. Gene sequencing, antigenic analysis and growth property were used to evaluate the rescued virus. Rescued virus show the genes sequence correct, keep the same antigenicity and similar growth property compared with wild type virus. The pandemic virus reverse genetics platform of A/California/07/2009 (H1N1) were built. Based on this platform, rescued virus hold the similarity of antigenicity and growth ability with wild type virus.


[Transfer of 45Ca during amelogenesis studied by autoradiography and electron microscopy].
45Ca transfer through the stratum intermedium and the secreting ameloblasts towards enamel has been studied by quantitative electron microscopical autoradiography in tooth-germs of newborn cats following intravenous injection of the isotope. Two transfer pathways were demonstrated. The relatively more important direct path passed through the stratum intermedium and ameloblast intercellular spaces and reached the enamel directly. The second pathway consisted in an intracellular transfer through the ameloblasts. 45Ca penetrated the cell through its basal pole. The mitochondria were the most highly labeled organelles at the different experimental time intervals studied. A maximum of radioactivity was respectively noticed at 30 min and 1 h in the endoplasmic reticulum and the Golgi apparatus. A total absence of silver grains was noted over the secretory ameloblastic bodies. At 6 h, the highest labeling was observed over enamel and especailly over the inner enamel along the enameldentin junction.


A mathematical model of T lymphocyte proliferation controlled by interleukin-2 internalization.
A mathematical model of the first in vitro IL-2-controlled division cycle of T lymphocytes is presented. The model describes the population dynamics of T cells at different stages of the cell cycle and introduces "molecular" equations for the G1-S phase control. These equations are based on the current knowledge of the biochemical mechanisms of ligand-receptor binding, ligand and receptor synthesis, and internalization of the ligand-receptor complexes. The temporal order in the biosynthesis of IL-2 and IL-2R during the G1a and G1b phases is investigated. We show numerically that the maximum number of cells in the S-phase depends on the reversibility of the G1a-G1b transitions, on the rates of ligand-receptor binding, ligand and receptor synthesis, and internalization of the ligand-receptor complexes. The phase portraits for different hypotheses on the temporal order of the IL-2 and high affinity receptor synthesis at the G1 phase of the cell cycle are qualitatively different. On this basis, simple kinetic experiments to distinguish between these alternatives are proposed.


Treatment of Malignant Pleural Mesothelioma: American Society of Clinical Oncology Clinical Practice Guideline.
Purpose To provide evidence-based recommendations to practicing physicians and others on the management of malignant pleural mesothelioma. Methods ASCO convened an Expert Panel of medical oncology, thoracic surgery, radiation oncology, pulmonary, pathology, imaging, and advocacy experts to conduct a literature search, which included systematic reviews, meta-analyses, randomized controlled trials, and prospective and retrospective comparative observational studies published from 1990 through 2017. Outcomes of interest included survival, disease-free or recurrence-free survival, and quality of life. Expert Panel members used available evidence and informal consensus to develop evidence-based guideline recommendations. Results The literature search identified 222 relevant studies to inform the evidence base for this guideline. Recommendations Evidence-based recommendations were developed for diagnosis, staging, chemotherapy, surgical cytoreduction, radiation therapy, and multimodality therapy in patients with malignant pleural mesothelioma. Additional information is available at www.asco.org/thoracic-cancer-guidelines and www.asco.org/guidelineswiki .


Impact of first contact on symptom onset-to-door time in patients presenting for primary percutaneous coronary intervention.
To determine effect of first medical contact type on symptom onset-to-door time (SODT). Shorter total ischemic time is associated with improved outcomes in ST-elevation myocardial infarction. From 2005 to 2009, we reviewed records of all consecutive patients treated with primary percutaneous coronary intervention for ST-elevation myocardial infarction at our tertiary care teaching hospital (median follow-up 3.85 years). We compared SODT in patients whose first medical contact was a private physician (in person or via telephone) vs patients who presented to the emergency department (ED) directly (in person or via Emergency Medical Services). Of 366 patients, 84 (23%) contacted a physician (group A) while 282 (77.6%) did not (group B). Group A had higher median SODT (239.5 vs 130 minutes, P = .0043) and significantly higher mortality (log rank P = .0392, Cox Proportional Hazard Model risk factors: physician contact first [P < .013], age [P < .0001] and peripheral vascular disease [P < .035]). Two factors associated with prolonged SODT: (1) contacting a physician first P = .002 and (2) personal mode of transportation, P = .002. Patients presenting during "on-hours" (weekdays) were more likely to first contact a physician compared with those presenting during "off-hours" (weeknights and weekends) (66.67% in group A vs 45.04% in group B, P < .001). Patients whose first medical contact was a physician had greater pre-hospital delays and worse survival compared to those who sought emergent medical care directly. This pattern occurred more often during "on-hours." Educational efforts aimed at both patient and physician office practices are warranted.


Reliability of Repeated Measurements on Post-Burn Scars with Corneometer CM 825(®).
The water content in burn scars, the parameter of stratum corneum water holding capacity, is an important feature in evaluation of biophysical properties of scars. Nevertheless, quantifiying this parameter is a challenge. In this study, the reliability of repeated water content measurements with Corneometer CM825(®) on (burn) scars was investigated. Intra-observer reliability, inter-observer reliability and day-by-day variability were examined on 30 scars by means of intra-class correlation coefficient (ICC) and within-subject coefficient of variation (WSCV). Bland-Altman plots with '95% limits of agreement' were constructed. Results revealed excellent ICC values (ICCintra = 0.985; ICCinter = 0.984) with relatively low WSCV (WSCVintra = 6.3%; WSCVinter = 10.6%) for respectively intra- and inter-observer reliability. However, the Bland-Altman plot showed that more than 5% of differences were expected to exceed 4 a.u., the limit of what has been defined as a clinically acceptable difference. Results for day-by-day variability showed good ICC value (ICCday-by-day = 0.849) and higher WSCV (WSCVday-by-day = 20.5%). The Corneometer CM825(®) is an objective and sensitive instrument for water content measurements. On the basis of our results, we concluded that the instrument can be used in clinical trials, but only under very strict conditions with standardized test protocol, preferably in combination with the evaluation of other physiological parameters.


Carbohydrate metabolism of malarial parasites--I. Metabolism of lactate in Plasmodium knowlesi infected monkey erythrocytes.
Lactate metabolism by Plasmodium knowlesi infected erythrocytes was examined after careful removal of leucocytes from cell preparations. Infected cells were able to metabolize glucose, pyruvate and lactate. Respiration of infected erythrocytes was maximally stimulated by lactate and to a lesser degree by pyruvate and glucose. Respiration of infected cells was insensitive to stimulation by succinate or glutamate or inhibition by malonate. Mepacrine was found to be a potent respiratory inhibitor. Chromatographic analysis of end products of lactate metabolism showed incorporation of carbon from [2-C14]lactate into phosphoenol pyruvate, 3-phosphoglycerate and malate. Experimental data failed to provide evidence for the presence of a functional citric acid cycle activity in infected cells.


Modeling of the Norwood circulation: effects of shunt size, vascular resistances, and heart rate.
Hypoplastic left heart syndrome is the most common lethal cardiac malformation of the newborn. Its treatment, apart from heart transplantation, is the Norwood operation. The initial procedure for this staged repair consists of reconstructing a circulation where a single outlet from the heart provides systemic perfusion and an interpositioning shunt contributes blood flow to the lungs. To better understand this unique physiology, a computational model of the Norwood circulation was constructed on the basis of compartmental analysis. Influences of shunt diameter, systemic and pulmonary vascular resistance, and heart rate on the cardiovascular dynamics and oxygenation were studied. Simulations showed that 1) larger shunts diverted an increased proportion of cardiac output to the lungs, away from systemic perfusion, resulting in poorer O2 delivery, 2) systemic vascular resistance exerted more effect on hemodynamics than pulmonary vascular resistance, 3) systemic arterial oxygenation was minimally influenced by heart rate changes, 4) there was a better correlation between venous O2 saturation and O2 delivery than between arterial O2 saturation and O2 delivery, and 5) a pulmonary-to-systemic blood flow ratio of 1 resulted in optimal O2 delivery in all physiological states and shunt sizes.


Inhibition of thyroid peroxidase by Myrcia uniflora flavonoids.
Thyroid peroxidase (TPO), the key enzyme in thyroid hormone biosynthesis, is inhibited by dietary flavonoids; thus, a high consumption of plants containing inhibitory flavonoids may affect thyroid function and lead to hypothyroidism. In this work, TPO inhibition by the aqueous partition of Myrcia uniflora and its isolated compounds has been evaluated. The aqueous partition of the methanolic extract of M. uniflora is able to inhibit TPO activity in vitro. Two known flavonoids were isolated and characterized by mass spectrometry and (1)H NMR from plant extracts: mearnsitrin and myricitrin. The degree of TPO inhibition produced by the aqueous solution of the flavonoids was very high, with a 50% inhibition of the original TPO activity (IC(50)) obtained at 1.97 microM mearnsitrin and at 2.88 microM myricitrin. These results suggest that the indiscriminated consumption of M. uniflora pharmaceutical products allied to the nutritional deficiency of iodine might contribute to the development of hypothyroidism and goiter.


Transport mechanisms of the large neutral amino acid L-phenylalanine in the human intestinal epithelial caco-2 cell line.
The transepithelial transport and the intracellular accumulation of the large neutral amino acid L-phenylalanine (L-Phe) were studied in monolayers of Caco-2 cells, cultivated in a bicameral insert system, to characterize the mechanisms involved in the absorption of this essential amino acid by the human intestinal mucosa. In our model, L-Phe was transported selectively in the apical (AP)-to-basolateral (BL) direction. AP-to-BL transport of L-Phe was temperature dependent and Na(+) independent, increased in the absence of protein synthesis and showed competition with large neutral and cationic amino acids. By contrast, transport in the BL-to-AP direction mainly resulted from passive movement (probably paracellular passage and transcellular diffusion). L-Phe accumulation into Caco-2 cells was higher from the BL pole than from the AP pole and characterized by the incorporation of most of the accumulated molecules into newly synthesized proteins. In addition, L-Phe accumulation was Na(+) dependent from both poles, whereas only accumulation from the AP pole was sensitive to inhibition by both large neutral and cationic amino acids. These results suggest that the processes involved in AP-to-BL transport and AP accumulation of this amino acid are very different from those involved in BL-to-AP transport and BL accumulation.


Juvenile homicide offenders: A 35-year-follow-up study.
Murders committed by juveniles have been a major concern in the USA for more than 40 years. Due to decisions in the 21st century by the United States Supreme Court, the likelihood of juvenile homicide offenders being sentenced to life without parole (LWOP) is less than in past decades. In addition, hundreds of individuals in prison who were sentenced to LWOP under mandatory sentencing statutes or their equivalent as juveniles for murder are now eligible for resentencing and may be released. Given these changes in sentencing policies and practices, follow-up research on juveniles involved in murder is needed. This study is a 35-year follow-up study of 59 boys who were convicted of murder and sentenced to adult prisons in a southeastern state, and initially interviewed in the early 1980s. Twenty-two of these men agreed to participate in clinical interviews about their experiences in prison and upon return to the community if released. The experiences in prison are highlighted for all men. Thereafter, attention focuses on the 19 juvenile homicide offenders who have been released. Of these 19 men, 11 were classified as failures given that they were sent back to prison at least once. Length of time served and completion of GED were the best predictors of post-release success. Clinical observations, implications of the findings, limitations of the study, and directions for future research are discussed.


Long-term effects of memory training in the elderly: a longitudinal study.
The effects of different types of memory training in young and older adults reported in a previous study [Cavallini, E., Pagnin, A., Vecchi, T., 2003. Aging and everyday memory: the beneficial effect of memory training. Arch. Gerontol. Geriatr. 37, 241-257] were again investigated from a longitudinal perspective 2 years after the original memory training sessions. The authors retested the original participants to measure the long-term effectiveness of two mnemonic strategies: the loci technique and strategic training. Three groups of participants (13 adults, M=24.1, 13 younger elderly, M=64.2 and 13 older elderly, M=74.4) were tested using a battery of seven tasks and four questionnaires, to evaluate memory performance and metamemory variables. The three age groups and the two trainings showed similar results on memory performance. Long-term effects were found only on two memory tasks, both were highly related to everyday life showing that, without additional practice, memory performance tended to go back to the original level. Moreover, the beneficial effects of the previous training sessions were particularly evident for older adults in metamemory knowledge and for strategic training in memory complaints. Results partially support the durability of memory training in improving memory performance.


Comparing Economics, Environmental Pollution and Health Efficiency in China.
As the modern economy develops rapidly, environmental pollution and human health have also been threatened. In recent years, relevant research has focused on subjects such as energy and economic, environmental pollution and health issues. Yet this has not considered the use of water resources and the impact of wastewater pollutant emissions on the economy and health. This article has combined the following factors like water consumption with wastewater discharge, pollutant concentration in sewage and local medical care expenditure and put them into the model of water resources, energy and health measurement, and a two-stage dynamic data envelopment analysis (DEA) model considering undesirable outputs is applied to 30 provinces (including autonomous regions and municipalities) to calculate the total efficiency, production efficiency and health efficiency in 2014-2017.The results show that the total efficiency values of most provinces are between 0.2 and 0.4, providing large room for improvement. Production efficiency and health efficiency have increased in recent years, but the health efficiency values of most provinces are still so low that they have dragged back the overall efficiency. The key impact indicators of different provinces are different, and each province should formulate different policies according to its own specific conditions so as to purposefully to deepen the energy, economic and medical reforms in each province, and also to promote sustainable economic development while improving health efficiency.


[Simultaneous determination of four aflatoxins in Citrus reticulatablanco by ultra performance liquid chromatography coupled with immunomagnetic beads for enrichment and purification].
Immunomagnetic beads (IMBs) were formed by coupling the ProtElut NHS (N-hydroxysuccinimide) magnetic beads and anti-aflatoxins monoclonal antibody. The synthesized IMBs presented uniform size, good magnetic property and specific selectivity. In this work, a novel and facile pretreatment method of sample enrichment and purification based on the IMBs for the determination of aflatoxins (AFs) (AFB1, AFB2, AFG1, AFG2) in Citrus reticulatablanco samples by ultra performance liquid chromatography (UPLC) were developed. The sample was extracted with 20% (v/v) methanol-PBS buffer solution (pH 7.4), followed by a cleanup procedure with the IMBs. The target compounds were eluted using 1 mL methanol. The four afatoxins were separated on an ACQUITY UPLC HSS T3 C18 column (100 mm x 2.1 mm, 1.8 μm) adopting a gradient program within 12 min. The UPLC equipped with a fluorescence detector with a special mercury/xenon lamp was used to detect the aflatoxins. The satisfactory correlation coefficients (r2 > 0.999) of the four aflatoxins were obtained within their respective linear ranges. The limits of detection (S/N = 3) were between 0.013 μg/kg and 0.038 μg/kg, and the limits of quantification (S/N = 10) were between 0.044 μg/kg and 1.2 μg/kg. The recoveries were in the range of 63.9% - 115.0% with the relative standard deviations (RSDs) of 0.4% - 14.2%. The validation results meet the requirements of trace assay. It's indicated that the IMBs were good pretreatment alternatives and the developed method is simple, fast, accurate, and can be applied for the simultaneous determination of the four aflatoxins in Citrus reticulatablanco.


Transit of radiopaque particles through the gastrointestinal tract: comparison between type 2 diabetes patients and healthy individuals.
some studies have reported controversial results when comparing the gastrointestinal transit between diabetic and healthy individuals. Therefore, we compared the gastrointestinal transit of radiopaque particles between diabetic and non-diabetic healthy individuals. abdominal radiographies were performed for 45 type 2 diabetes mellitus patients and 35 healthy individuals (gender and age similar for both groups) at 24 and 72 h after they ingested radiopaque particles. The mean number of particles in the colon was compared for both groups. The data were expressed as mean and standard deviation values. at 24 h, the total number of particles in the colon did not differ significantly for the diabetic and non-diabetic individuals. At 72 hours, the distribution in the diabetic and non-diabetic individuals was as follows: right colon, 0.44 ± 0.88 and 0.26 ± 0.7, respectively (p = 0.8); left colon, 2.6 ± 4.2 and 0.49 ± 1.3 (p < 0.003); and rectosigmoid colon, 2.65 ± 3.8 and 0.80 ± 1.5 (p < 0.005).The mean number of radiopaque particles in the entire colon was 5.7 ± 7.1 and 1.5 ± 2.7 for diabetic and non-diabetic individuals, respectively (p < 0.001). the number of radiopaque particles in the colon did not significantly differ for the diabetic and non-diabetic individuals at 24 h after ingestion but was significantly greater in diabetic individuals at 72 h after ingestion. At 72 h, the mean number of radiopaque particles in the left and rectosigmoid colon were significantly higher in the diabetics than in the non-diabetic individuals.


Forty-year survival with Smeloff-Cutter and Starr-Edwards prostheses.
This case study describes a 40-year follow-up of a man who had a double valve replacement with Smeloff-Cutter aortic (Cutter Laboratories, Berkeley, CA) and Starr-Edwards mitral prostheses (Edwards Laboratories, Santa Ana, CA) when he was 34 years old. Double valve replacement was performed for aortic and mitral valve stenosis and insufficiency. To date, no surgical treatment has been required except a pacemaker implantation. The patient presented with a New York Heart Association functional class of I to II. Echocardiography revealed intact prostheses.


Vasodilating actions of cromakalim in resting and contracting states of carotid arteries from spontaneously hypertensive rats.
To determine the properties of cromakalim-opened K+ channels in arterial smooth muscle of spontaneously hypertensive rats (SHR), the effects of cromakalim on tension and 86Rb efflux were compared in endothelium-denuded strips of carotid arteries from 13-week-old SHR and normotensive Wistar-Kyoto rats (WKY). The addition of cromakalim or of nifedipine to resting strips caused a relaxation only in SHR. When strips from both strains were contracted with 15.9 mM K+, the magnitude of the precontraction was greater in SHR than in WKY. Under these conditions, relaxant responses to lower concentrations of cromakalim were decreased and those to higher concentrations of cromakalim were increased in SHR. When strips from both strains were contracted with a different concentration of K+ to an equivalent magnitude (78% of the maximum) relaxant responses to cromakalim were greater in SHR than in WKY. When strips were contracted with 10(-7) M norepinephrine, the precontraction was similar between SHR and WKY, and relaxant responses to cromakalim were greater in SHR. In both strains, the relaxant responses to cromakalim were competitively antagonized by glibenclamide, a blocker of ATP-sensitive K+ (KATP) channels, with a pA2 value of approximately 7.3. Charybdotoxin (10(-7) M), a blocker of Ca(2+)-activated K+ (KCa) channels, did not inhibit the relaxant responses to cromakalim in both strains. Charybdotoxin alone elicited a contraction, which was greater in SHR than in WKY. In resting strips preloaded with 86Rb, the basal 86Rb efflux rate constant was higher in SHR than in WKY. The addition of cromakalim (10(-5) M) to the resting strips increased the 86Rb efflux rate constant in both strains to a similar peak value. The addition of nifedipine (10(-7) M) to the resting strips decreased the basal 86Rb efflux rate constant only in SHR, and concomitantly affected the action of cromakalim in SHR. The results suggest that (1) cromakalim caused arterial relaxation via the opening of KATP channels in both SHR and WKY, (2) although the relaxant effects of cromakalim tended to be greater in SHR than in WKY, the differences were rather small, depended on the precontraction tone and varied with the concentration of the vasoconstrictors, and (3) there was an increased basal Ca2+ influx and a high activation of KCa channels in the resting state of SHR arteries, and these changes might influence the effects of cromakalim.


Human retinal pigment epithelial cells possess muscarinic receptors coupled to calcium mobilization.
Human retinal pigment epithelial (RPE) cells in culture demonstrated saturable specific binding of [3H]quinuclidinyl benzilate (QNB). Specific binding represents about 75% of total binding. Scatchard analysis yields a Kd of 0.178 nM and Bmax of 42 fmol/mg protein. Atropine and carbachol show typical displacement curves, and a Hill plot has a slope of 0.96, suggesting a homogeneous population of receptors. Muscarinic agonists have no effect on intracellular cyclic adenosine monophosphate levels in RPE cells measured by radioimmunoassay, nor do they alter the isoproterenol-induced stimulation of adenylate cyclase. However, both acetylcholine and carbachol cause a rapid increase in intracellular calcium concentration measured by the fluorescent indicator quin 2. Atropine reverses the calcium rise when added after agonist and prevents the rise when added prior to agonist. These data suggest that human RPE cells possess muscarinic receptors coupled to calcium mobilization.


Tight junctions in epithelial cells of human fetal hindgut, normal colon, and colon adenocarcinoma.
The structural patterns of tight junctions in normal human colon mucosa, colon adenocarcinomas, and fetal colon were studied and compared by the freeze-fracturing technique. The zonula occludens of the normal colon cells at the upper, more differentiated part of the crypts of Lieberkühn appeared as continuous belts made of about eight parallel strands. At the less differentiated bases of the crypts, the zonula occludens was less regular and contained fewer, mostly beaded strands. In the colons of 10-week fetuses, early stages of tight junction assembly were observed. At the same time, vesicles bearing remnants of tight junction elements were observed within the cytoplasm. This finding suggested that during the early development and organization of the fetal gut, mechanisms of assembly and disassembly of tight junctions are operating concomitantly. In well-differentiated adenocarcinomas, the cells in the luminal region retained their polarity and had seven or eight parallel junctional elements. In infiltrating cells, however, tight junctions appeared as fascia occludens and resembled the junctional organization of 10-week fetuses.


Organic carbon effects on aerobic polychlorinated biphenyl removal and bacterial community composition in soils and sediments.
Certain organic compounds, including biphenyl and salicylic acid, stimulate polychlorinated biphenyl (PCB) degradation by microorganisms in some environments. However, the usefulness of these amendments for improving PCB removal by microorganisms from diverse habitats has not been extensively explored. This study evaluated the effects of biphenyl, salicylic acid, and glucose on changes in aerobic PCB removal and bacterial communities from an agricultural soil, a wetland peat soil, a river sediment, and a mixture of these samples. PCB removal patterns were significantly different between soils and sediments amended with carbon compounds: (i) terrestrial soil microorganisms removed more PCBs than river sediment microorganisms, particularly with regard to PCBs with >4 chlorine substituents, (ii) glucose-supplemented, agricultural soil microorganisms removed more hexachlorobiphenyl than unsupplemented samples, (iii) biphenyl-supplemented, river sediment microorganisms removed more di- and tri-chlorobiphenyls than unamended samples. Carbon amendments also caused unique shifts in soil and sediment bacterial communities, as determined by specific changes in bacterial 16S rRNA denaturing gradient gel electrophoresis banding patterns. These results indicate that organic carbon amendments had site-specific effects on bacterial populations and PCB removal. Further work is needed to more accurately characterize PCB degrading communities and functional gene expression in diverse types of environments to better understand how they respond to bioremediation treatments.


Sudden unexpected death in infants of narcotic-dependent mothers.
During the study years 1972--1974, 8 of 383 infants born to mothers with known narcotic dependency during pregnancy died unexpectedly within the first 4 mth of life; autopsies were compatible with the diagnosis of SIDS. This incidence of SIDS was 5.5 times that in our hospital populations (P < 0.001) and 8.7 times that of our borough within New York City (P < 0.001). Similar factors, such as sex ratio, age at time of death, and diurnal and seasonal variations suggest that narcotic-associated sudden death may be a relevant study model for sudden unexpected death in the general population. Intrauterine exposure to narcotics and its subsequent effect on central control of respiration in the young infant may be the underlying mechanism of drug related SIDS.


The informed consent aftermath of the genetic revolution. An Italian example of implementation.
A great part of human genetics research is carried out collecting data and building large databases of biological samples that are in a non-anonymous format. These constitute a valuable resource for future research. The construction of such databases and tissue banks facilitates important scientific progress. However, biobanks have been recognized as ethically problematic because they contain thousands of data that could expose individuals and populations to discrimination, stigmatization and psychological stress if misused. Informed consent is regarded as a cornerstone in the protection of personal autonomy in research involving human subjects. Yet in recent years this fundamental concept has been overwhelmed by the genomic revolution. From a general overview of international literature, it seems evident that informed consent issues have come into sharp focus, in particular in relation to the twin issues of time extension (blanket versus specific/repeated consent) and personal extension (group consent). After an introduction on obtaining informed consent in the context of genetic research, this paper addresses the apparent lack of a single, universal model of obtaining informed consent among populations involved in genetic research and it argues for the need to develop an ethical framework tailored to the specific features of each project. In order to support this theory of contextualizing, the case of a private biotechnology company, SharDNA is presented. The present paper explores the management of its biobank, developed from a genetic research project carried out on isolated populations living on the Italian island of Sardinia. In particular, the paper highlights how the company is tackling the problem of informed consent and other ethical requirements for genetic research, such as the respect of individual privacy, the population approach and the existing Italian legal regulatory framework.


Role of soil-to-leaf tritium transfer in controlling leaf tritium dynamics: Comparison of experimental garden and tritium-transfer model results.
Environmental transfer models assume that organically-bound tritium (OBT) is formed directly from tissue-free water tritium (TFWT) in environmental compartments. Nevertheless, studies in the literature have shown that measured OBT/HTO ratios in environmental samples are variable and generally higher than expected. The importance of soil-to-leaf HTO transfer pathway in controlling the leaf tritium dynamics is not well understood. A model inter-comparison of two tritium transfer models (CTEM-CLASS-TT and SOLVEG-II) was carried out with measured environmental samples from an experimental garden plot set up next to a tritium-processing facility. The garden plot received one of three different irrigation treatments - no external irrigation, irrigation with low tritium water and irrigation with high tritium water. The contrast between the results obtained with the different irrigation treatments provided insights into the impact of soil-to-leaf HTO transfer on the leaf tritium dynamics. Concentrations of TFWT and OBT in the garden plots that were not irrigated or irrigated with low tritium water were variable, responding to the arrival of the HTO-plume from the tritium-processing facility. In contrast, for the plants irrigated with high tritium water, the TFWT concentration remained elevated during the entire experimental period due to a continuous source of high HTO in the soil. Calculated concentrations of OBT in the leaves showed an initial increase followed by quasi-equilibration with the TFWT concentration. In this quasi-equilibrium state, concentrations of OBT remained elevated and unchanged despite the arrivals of the plume. These results from the model inter-comparison demonstrate that soil-to-leaf HTO transfer significantly affects tritium dynamics in leaves and thereby OBT/HTO ratio in the leaf regardless of the atmospheric HTO concentration, only if there is elevated HTO concentrations in the soil. The results of this work indicate that assessment models should be refined to consider the importance of soil-to-leaf HTO transfer to ensure that dose estimates are accurate and conservative.


Proteins without 3D structure: definition, detection and beyond.
Predictions, and experiments to a lesser extent, following the decoding of the human genome showed that a significant fraction of gene products do not have well-defined 3D structures. While the presence of structured domains traditionally suggested function, it was not clear what the absence of structure implied. These and many other findings initiated the extensive theoretical and experimental research into these types of proteins, commonly known as intrinsically disordered proteins (IDPs). Crucial to understanding IDPs is the evaluation of structural predictors based on different principles and trained on various datasets, which is currently the subject of active research. The view is emerging that structural disorder can be considered as a separate structural category and not simply as absence of secondary and/or tertiary structure. IDPs perform essential functions and their improper functioning is responsible for human diseases such as neurodegenerative disorders.


SUMOylation and de-SUMOylation in response to DNA damage.
To maintain genomic integrity, a cell must utilize multiple mechanisms to protect its DNA from the damage generated by environmental agents or DNA metabolism. SUMO (small ubiquitin-like modifier) can regulate protein stability, protein cellular location, and protein-protein interactions. In this review, we summarize the current understanding of the roles of SUMOylation and de-SUMOylation in DNA damage response (DDR) and DNA repair with a specific focus on the role of RPA SUMOylation in homologous recombination (HR).


Estimation of inertial parameters of the lower trunk in pregnant Japanese women: A longitudinal comparative study and application to motion analysis.
We aimed to quantify the inertial parameters of the lower trunk segment in pregnant Japanese women and compare kinetic data during tasks calculated with parameters estimated in this study to data calculated with standard parameters. Eight pregnant women and seven nulliparous women participated. Twenty-four infrared reflective markers were attached to the lower trunk, and the standing position was captured by eight infrared cameras. The lower trunk was divided into parts, and inertial parameters were calculated. Pregnant women performed a movement task that involved standing from a chair, picking up plates, and walking forward after turning to the right. Kinetic analysis was performed using standard inertial parameters and the newly calculated parameters. There were more significant differences between methods in the kinetic data at the latter stages of pregnancy. The inertial parameters calculated in this study should be used to ensure the validity of biomechanical studies of pregnant Japanese women.


Morphometric changes in the acetabulum after Dega osteotomy in patients with cerebral palsy.
We carried out a morphometric analysis of the acetabulum following Dega osteotomy in patients with cerebral palsy using three-dimensional CT. We assessed 17 acetabula in 12 patients with instability of the hip. A Dega osteotomy and varus derotation femoral osteotomy were performed in all 17 hips. Three-dimensional CT scans were taken before and approximately one year after operation. Acetabular cover was evaluated using anterosuperior, superolateral and posterosuperior acetabular indices, and the change in the acetabular volume was calculated. Inter- and intra-observer reliability was assessed using the intraclass correlation coefficient. After the osteotomy, the anterosuperior, superolateral and posterosuperior cover had improved significantly towards the value seen in a control group. The mean acetabular volume increased by 68%.


Linking social and ecological systems to sustain coral reef fisheries.
The ecosystem goods and services provided by coral reefs are critical to the social and economic welfare of hundreds of millions of people, overwhelmingly in developing countries [1]. Widespread reef degradation is severely eroding these goods and services, but the socioeconomic factors shaping the ways that societies use coral reefs are poorly understood [2]. We examine relationships between human population density, a multidimensional index of socioeconomic development, reef complexity, and the condition of coral reef fish populations in five countries across the Indian Ocean. In fished sites, fish biomass was negatively related to human population density, but it was best explained by reef complexity and a U-shaped relationship with socioeconomic development. The biomass of reef fishes was four times lower at locations with intermediate levels of economic development than at locations with both low and high development. In contrast, average biomass inside fishery closures was three times higher than in fished sites and was not associated with socioeconomic development. Sustaining coral reef fisheries requires an integrated approach that uses tools such as protected areas to quickly build reef resources while also building capacities and capital in societies over longer time frames to address the complex underlying causes of reef degradation.


Clonality structure in Candida dubliniensis.
Multilocus enzyme electrophoresis was performed on 76 European strains of Candida dubliniensis. Ten of the 20 enzyme-encoding loci were polymorphic, giving rise to 10 electrophoretic types within the sample studied. Investigation of the population genetics of a subset of 36 strains from HIV-infected patients in London showed the existence of strong heterozygote deficits and excesses associated with significant linkage disequilibria between pairs of loci. These findings, together with the predominance of multilocus genotypes, strongly suggest that C. dubliniensis is mainly (if not totally) clonal. Analysis of genotypes of a larger number of strains should confirm this conclusion and improve our understanding of the epidemiology of this pathogen.


Cardiovascular diseases and genome-wide association studies.
Genome-Wide Association Studies (GWAS) on cardiovascular diseases and related quantitative traits revealed numerous genetic variants, which however have been partially replicated, probably due to the heterogeneity of the clinical phenotypes and the populations studied. Even if novel biological pathways have been identified through these studies, there is still a long way until the validation of causal variants and their use in clinical practice as factors for prevention, risk assessment and as targets for the development of new medications. GWAS methodologies should, in the following years, integrate gene-gene and gene-environment interaction analyses in a global research strategy and also involve subsequent transcriptomic and proteomic investigations. The GWAS era is very promising but it is just at the beginning.


Targeting apoptosis via chemical design: inhibition of bid-induced cell death by small organic molecules.
Bid is a key member of the Bcl-2 family proteins involved in the control of the apoptotic cascade in cells, leading to cell death. Uncontrolled cell death is associated with several human pathologies, such as neurodegenerative diseases and ischemic injuries. Therefore, Bid represents a potential yet unexplored and challenging target for strategies aimed at the development of therapeutic agents. Here we show that a multidisciplinary NMR-based approach that we named SAR by ILOEs (structure activity relationships by interligand nuclear Overhauser effect) allowed us to rationally design a series of 4-phenylsulfanyl-phenylamine derivatives that are capable of occupying a deep hydrophobic crevice on the surface of Bid. These compounds represent the first antiapoptotic small molecules targeting a Bcl-2 protein as shown by their ability to inhibit tBid-induced SMAC release, caspase-3 activation, and cell death.


NEMO trimerizes through its coiled-coil C-terminal domain.
NEMO/IkappaB kinase (IKK) gamma is the regulatory component of the IKK complex comprising the two protein kinases, IKKalpha and IKKbeta. To investigate the self-assembly properties of NEMO and to understand further the mechanism of activation of the IKK complex, we purified wild-type and mutant NEMO expressed in Escherichia coli. In the absence of its IKK partners, recombinant NEMO (rNEMO) is a metastable functional monomer correctly folded, according to its fluorescence and far-UV CD spectra, which is binding specifically to the IKK complex. A minor fraction of rNEMO was found tightly associated with DnaK (E. coli Hsp70). We also examined the interaction of NEMO with prokaryotic and eukaryotic Hsp70, and we showed that the Hsp70-NEMO complex forms a supramolecular structure probably corresponding to an assembly intermediate. In vivo cross-linking experiments indicate that native NEMO in association with IKK is in equilibrium between a dimeric and a trimeric form. Similarly to native NEMO, a NEMO mutant deleted from its IKK binding N-terminal domain (residues 242-388) forms a stable trimeric coiled-coil, suggesting that the association of NEMO with IKK or with Hsp70 prevents incorrect interdomain pairing reactions that could lead to aggregation or to an non-native oligomeric state of rNEMO. We propose a model in which the activation of the IKK complex occurs through the trimerization of NEMO upon binding to a not yet identified upstream activator.


Investigating a Multistakeholder Alliance Approach to Reducing Hospital Readmissions.
There is growing appreciation that significant improvements in healthcare quality cannot be achieved by organizations working in isolation. To that end, the Robert Wood Johnson Foundation's Aligning Forces for Quality (AF4Q) initiative provides multistakeholder alliances-groups of payers, purchasers, providers, and consumers that work together-with funding and technical assistance to improve healthcare community wide. Reducing readmissions was one of the several AF4Q goals. Alliances crafted their own interventions, tailored to address the communities' unique needs, and took advantage of local resources. Our objective was to determine whether readmission rates declined more in AF4Q communities than in non-AF4Q communities by year 7 of the 10-year initiative. We conducted a quasiexperimental study using zip code-level demographics and 30-day readmission rates for all medical and surgical discharges using Medicare claims data for 2008 through 2012. This interim analysis found that AF4Q was not associated with significant reductions in medical or surgical readmissions, suggesting that AF4Q's multistakeholder approach was no more effective at reducing readmissions than other national efforts, such as the Hospital Reduction Program under the Affordable Care Act.


Disappearance of human chorionic gonadotropin and resumption of ovulation following abortion.
The disappearance of human chorionic gonadotropin (hCG) and resumption of pituitary ovarian function was investigated in 13 patients following first- and second-trimester abortions. First-trimester abortion patients (with suction curettage) had a mean time of 37.5 +/- 6.4 days for the clearance of hCG to a level of 2 mlU/ml. Second trimester abortions (with prostaglandin) had a mean time of 27.4 +/- 4.8 days. Patients undergoing second-trimester hysterectomy had a mean disappearance time of 39.7 +/- 5.3 days and only 12 days if the hysterectomy was initiated with ligation of the uterine and ovarian vessels. No significant difference in clearance time was found when it was compared on the basis of the baseline hCG levels. Based on a concomitant luteinizing hormone (LH) and follicle-stimulating hormone (FSH) peak, nine of 12 patients resumed normal pituitary function. These LH and FSH peaks were seen even though the serum hCG levels were as high as 35 mlU/ml. Based on serum progesterone levels of greater than 3 ng/ml, all these nine patients ovulated as early as 21 days after abortion. In view of these results, the clearance of hCG after pregnancy termination depends mainly upon the type of procedure used. Moreover, in view of the early time of ovulatory recovery, contraception should be instituted within the first 2 weeks following pregnancy termination.


Carbonate-Promoted Drift of Alkali Cations in Small Pore Zeolites: Ab Initio Molecular Dynamics Study of CO2 in NaKA Zeolite.
An effect of deblocking of small size (8R, D8R) pores in zeolites due to cation drift is analyzed by using ab initio molecular dynamics (AIMD) at the PBE-D2/PAW level. The effect of carbonate and hydrocarbonate species on the carbon dioxide uptake in NaKA zeolite is demonstrated. It is shown that a hydrocarbonate or carbonate anion can form strong complexes with K+ cation and withdraw it from the 8R window, so that the probability of CO2 diffusion through 8R increases. For the first time, correlations between cationic and HCO3-/CO32- positions are demonstrated in favor of their significant interaction leading to the cationic drift from 8R windows. This phenomenon explains a nonzero CO2 adsorption in narrow pore zeolites upon high Na/K exchange. In a gas mixture, such deblocking effect reduces the separation factor because of the possible passage of both components through the plane of partly open 8R windows.


Multiple molecular forms of prostaglandin 15-hydroxydehydrogenase and 9-ketoreductase in chicken kidney.
Prostaglandin-15-hydroxydehydrogenase and prostaglandin-9-keto-reductase were purified from chicken kidney. Both enzymes exist in multiple forms as determined by isoelectric focusing. The dehydrogenases catalyze the transformation of the functional group at C-15 but not the functional group at C-9. The preferred cofactors in these reactions are NAD+ or NADH. The 9-ketoreductases catalyze the reversible transformation of the functional group at C-9 and also the oxidation or reduction of the C-15 functional group. The preferred cofactors are NADP+ or NADPH. Bradykinin does not affect the activities of any of the three prostaglandin 9-ketoreductases. Flavin mononucleotide and the flavonoid, quercetin, as well as indomethacin, ethacrynic acid, and furosemide, inhibit all three 9-ketoreductases. An inhibitor of 9-ketoreductase isolated from chicken breast muscle also inhibits the three separable reductases, but the pattern of inhibition of the reductase that focuses at pH 5.7 differs from that of the reductases focusing at pH 7.8 and 8.2.


Possible isozyme-specific effects of experimental malaria infection with Plasmodium berghei on cytochrome P450 activity in rat liver microsomes.
We have investigated the effect of experimental malaria infection on rat cytochrome P450-mediated drug metabolism using ethoxyresorufin and metoprolol as probe compounds. Malaria infection caused a significant reduction in total intrinsic clearance of ethoxyresorufin in both low and high parasitaemia malaria compared with control (control 18.7 +/- 7.2; low parasitaemia 10.5 +/- 4.1; high parasitaemia 4.3 +/- 1.4 mL min-1). However, clearance of metoprolol was unchanged in malaria infection compared with control (control 2.7 +/- 1.2; malaria 4.0 +/- 1.7 mL min-1). The change in clearance of ethoxyresorufin was the result of a decrease in Vmax, with no apparent change in Km. There was no change in either Vmax or Km of metoprolol. These results indicate a possible isozyme-selective effect of experimental malaria.


[Perinatal bereavement and support for siblings].
When the premature death of a baby occurs in a family, brothers and sisters need support. Their silence, their absence of questions or reactions must set off alarm bells. The parents, beyond their own bereavement, have a key role to play in enabling their children to integrate this loss into their lives.


Dose-response relationship of sertindole and haloperidol using the pharmacopsychometric triangle.
Renewed insight into dose-related effects of sertindole and haloperidol was sought by re-analysing published data for antipsychotic effect, extrapyramidal effect, and patient wellbeing - i.e., the important pharmacopsychometric triangle domains. Selected Positive and Negative Syndrome Scale (PANSS) subscales and the Simpson-Angus scale were tested for validity. Standardized effect sizes [last observation carried forward (LOCF)] at endpoint were calculated. The scales were found to be valid instruments. The PANSS(11) psychotic subscale showed clinically significant effect sizes for all doses of sertindole (12, 20, and 24 mg) and haloperidol (4, 8, and 16 mg). Extrapyramidal effects were evident for all doses of haloperidol, but absent for the lower doses of sertindole. The PANSS(6) depression subscale, a proxy measure of quality of life, showed a clinically significant effect for sertindole 20 mg and no effect for haloperidol. This re-analysis confirmed the antipsychotic effect and absence of extrapyramidal effects for sertindole and, in addition, showed a clinically significant antidepressant effect. A profile for bipolar states emerged.


Scatter/primary ratios for x-ray spectra modified to enhance iodine contrast in screen-film mammography.
Contrast mammography to detect the uptake of iodine-containing contrast material may be enhanced by spectral modification of the x-ray beam. Luminance scatter-to-primary ratios were measured for three candidate x-ray tube anode/filter combinations (Mo/Mo, W/Ce, and Ce/Ce). Results show that scattered radiation is significant for all tubes, is lowest for the Mo/Mo system and is essentially the same for the tungsten and cerium anode systems.


Multiple mounting and staining on 2" x 2" glass slides.
A technique is described for multiple mounting and staining of histopathologic material on 2'' times 2'' glass slides. Various applications as well as the advantages and disadvantages of this procedure are discussed.


Effects of intracameral injection of preservative-free lidocaine on the anterior segment of the eyes in dogs.
To evaluate effects of intracameral injection of preservative-free 1% and 2% lidocaine hydrochloride solution on the anterior segment of the eyes in dogs. 16 adult healthy dogs (8 male and 8 female) judged to be free of ocular disease. Dogs were randomly assigned to 2 groups of 8 dogs each. Group 1 dogs received an intracameral injection of 0.10 mL of preservative-free 1% lidocaine solution in the designated eye, and group 2 dogs received 0.10 mL of preservative-free 2% lidocaine solution in the designated eye. After injection, intraocular pressure was measured every 12 hours for 48 hours and then every 24 hours until 168 hours after injection. Slit-lamp biomicroscopy was performed preceding intracameral injection, 8 hours after injection, and then every 24 hours until 168 hours after injection. Ultrasonic pachymetry and specular microscopy were performed preceding intracameral injection and 72 and 168 hours after injection. Corneal thickness and endothelial cell density and morphology were compared with baseline measurements. No significant differences were found in intraocular pressure, corneal thickness, endothelial cell density, and morphologic features in either group, compared with baseline. A significant difference in aqueous flare was found for treated and control eyes 8, 24, and 48 hours after injection, compared with baseline. No significant difference in aqueous flare was found between treated and control eyes within either group. No adverse ocular effects were detected after intracameral injection of preservative-free 1% or 2% lidocaine solution; thus, its use would be safe for intraocular pain management in dogs.


[Diagnosis of diabetic macroangiopathy of the lower extremities using rheography].
Rheography (impedance plethysmography) makes it possible to differentiate the normal condition of arteries of the lower extremity from functional and organic affections. The examination of a group of 325 patients with diabetes mellitus revealed that without the use of rheography, i.e. when relying only on examination of the lower extremities by palpation, the presence of macroangiopathy would escape attention in 46% of the affected subjects. Macroangiopathy of an organic character was found significantly more frequently in type 2 (NIDDM) (35%) than in type 1 (IDDM) (9%; p less than 0.03). Its incidence increased in both types of diabetes with the duration of the disease. Moreover it was significantly associated with hyperlipoproteinaemia. The assembled results drew attention to the great diagnostic value of rheography. It permits also to document objectively the effect of treatment focused on macroangiopathy.


Culture of bovine oviduct epithelial cells (BOEC).
Bovine oviduct epithelial cells are widely used in co-culture experiments to improve early embryonic development and in vitro fertilization in embryo transfer programmes for domestic animals. The present study compares different methods for harvesting and culture of bovine oviduct epithelial cells in order to optimize handling. Bovine oviduct epithelial cells were mechanically or enzymatically isolated and cultured on glass, on permeable membranes, or in suspension. Growth of the cells and their state of differentiation was examined by means of classical staining methods, immunohistochemistry and electron microscopy. Initial cell suspensions contained sheets of ciliated and nonciliated (secretory) cells; 24 h after seeding, free floating epithelial cells formed vesicles with cilia on their external surface. First adhesion of cells was seen 72 h after seeding. Later on, cells grew continuously and confluent monolayers were formed after 7 days. Results were identical after mechanical or enzymatical cell harvesting and were identical on both substrata tested, i.e., on glass and on permeable membranes. Light and electron microscopy proved the monolayers to resemble a polarized, simple, cuboidal to columnar epithelial membrane with intact junctional complexes and numerous apical microvilli. Their epithelial nature was established by immunostaining for cytokeratins. Cilia were missing and secretory granules were scarce. A layer of acidic glycoprotein material was demonstrated on the apical surface. Monolayers of bovine oviduct epithelial cells stored lipid droplets and large quantities of glycogen. About 50% of the seeded cells did not adhere but survived in the culture medium as free floating cells. These suspended cells maintained morphological criteria of differentiation (cilia and secretory granules) until day 12 of culture. Proliferation rates of cultivated cells were determined by counting mitoses and by immunostaining with MIB1 antibody. Results showed coincidence of rapid proliferation and morphological dedifferentiation of monolayers. Suspended cells, by contrast, did not proliferate but retained cellular differentiation under identical culture conditions. The results strongly suggest that monolayers of bovine oviduct epithelial cells will not fully substitute for original oviduct epithelium when used in co-culture experiments after in vitro fertilization.


Leg glucose and protein metabolism during an acute bout of resistance exercise in humans.
The present study investigated the responses of leg glucose and protein metabolism during an acute bout of resistance exercise. Seven subjects (5 men, 2 women) were studied at rest and during a strenuous lower body resistance exercise regimen consisting of approximately 8 sets of 10 repetitions of leg press at approximately 75% 1 repetition maximum and 8 sets of 8 repetitions of knee extensions at approximately 80% 1 repetition maximum. L-[ring-2H5]phenylalanine was infused throughout the study for measurement of phenylalanine rates of appearance, disappearance, protein synthesis, and protein breakdown across the leg. Femoral arterial and venous blood samples were collected at rest and during exercise for determination of leg blood flow, concentrations of glucose, lactate, alanine, glutamine, glutamate, leucine, and phenylalanine, and phenylalanine enrichments. Muscle biopsies were obtained at rest and immediately after exercise. Leg blood flow was nearly three times (P <0.009) higher and glucose uptake more than five times higher (P=0.009) during exercise than at rest. Leg lactate release was 86 times higher than rest during the exercise bout. Although whole body phenylalanine rate of appearance, an indicator of whole body protein breakdown, was reduced during exercise; leg phenylalanine rate of appearance, rate of disappearance, protein synthesis, and protein breakdown did not change. Arterial and venous alanine concentrations and glutamate uptake were significantly higher during exercise than at rest. We conclude that lower body resistance exercise potently stimulates leg glucose uptake and lactate release. In addition, muscle protein synthesis is not elevated during a bout of resistance exercise.


Lipodystrophy and metabolic abnormalities in a case of adult dermatomyositis.
Lipodystrophy and metabolic abnormalities, primarily hypertriglyceridemia and insulin resistance, have been reported in juvenile dermatomyositis. We report a 55-year-old woman with adult dermatomyositis who developed lipodystrophy of the thighs, hypertriglyceridemia, and insulin resistance. Our case illustrates that lipodystrophy may occur in adult and juvenile dermatomyositis. Loss of subcutaneous tissue may be a cutaneous marker for metabolic abnormalities in both the adult and the juvenile forms of dermatomyositis.


Self-assembling peptide nanostructures on aligned poly(lactide-co-glycolide) nanofibers for the functional regeneration of sciatic nerve.
Regeneration of functional peripheral nerve tissue at critical-sized defect requires extracellular matrix analogs impregnated with appropriate biosignals to regulate the cell fate process and subsequent tissue progression. The aim of the study was to develop electrospun aligned nanofibers as architectural analogs integrated with RADA16-I-BMHP1 as biofunctional peptides. Aligned poly(lactide-co-glycolide) (PLGA)-RADA16-I-BMHP1 nanofibers were fabricated and characterized for their in vitro potential using rat Schwann cell line and in vivo potential using a 10 mm sciatic nerve transection rat model. PLGA-peptide scaffolds significantly promoted higher expression of genotypic markers and bipolar extension of Schwann cells. Further, PLGA-peptide treated animals promoted the native collagen organization, remyelination and showed significantly higher recovery of sensorimotor and motor function than PLGA-treated groups (p < 0.05). Our results demonstrate that self-assembling peptide nanostructures on aligned PLGA nanofibers provided better cell-matrix communication with significant functional restoration of the sciatic nerve.


The effect of dimerization on the activation and conformational dynamics of adenosine A1 receptor.
The adenosine A1 receptor (A1R) is one of four adenosine receptors in humans, which are involved in the function of the cardiovascular, respiratory and central nervous systems. Experimental results indicate that A1R can form a homodimer and that the protomer-protomer interaction in the A1R dimer is related to certain pharmacological characteristics of A1R activation. In this work, we performed docking, metadynamics simulation, conventional molecular dynamics simulations, Gaussian-accelerated molecular dynamics simulations, potential of mean force calculations, dynamic cross-correlation motions analysis and community network analysis to study the binding mode of 5'-N-ethylcarboxamidoadenosine (NECA) to A1R and the effect of dimerization on the activation of A1R. Our results show that NECA binds to A1R in a similar mode to adenosine in the A1R crystal structure and NECA in the A2AR crystal structure. The A1R homodimer can be activated by one or two agonists with NECA occupying its orthosteric pockets in one (which we call the NECA-A1R system) or both protomers (which we call the dNECA-A1R system). In the NECA-A1R system, activation is predicated in the protomer without NECA bound. In the dNECA-A1R system, only one protomer achieves the active state. These findings suggest an asymmetrical activation mechanism of the homodimer and a negative cooperativity between the two protomers. We envision that our results may further facilitate the drug development of A1R.


Recommendations for the validation of cell-based assays used for the detection of neutralizing antibody immune responses elicited against biological therapeutics.
The administration of biological therapeutics may result in the development of anti-drug antibodies (ADAs) in treated subjects. In some cases, ADA responses may result in the loss of therapeutic efficacy due to the formation of neutralizing ADAs (NAbs). An important characteristic of anti-drug NAbs is their direct inhibitory effect on the pharmacological activity of the therapeutic. Neutralizing antibody responses are of particular concern for biologic products with an endogenous homolog whose activity can be potentially dampened or completely inhibited by the NAbs leading to an autoimmune-type deficiency syndrome. Therefore, it is important that ADAs are detected and characterized appropriately using sensitive and reliable methods. The design, development and optimization of cell-based assays used for detection of NAbs have been published previously by Gupta et al. 2007 [1]. This paper provides recommendations on best practices for the validation of cell-based NAb assay and suggested validation parameters based on the experience of the authors.