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Focal adhesion kinase (FAK) contributes to cellular homeostasis under stress conditions. Here, we show that αB-crystallin confers protection to FAK against calpain-mediated proteolysis under mechanical stress in cardiomyocytes. Biochemical assays, chemical cross-linking coupled to mass spectrometry experiments, mutational analyses and Förster resonance energy transfer (FRET) were combined to investigate the basis of FAK and αB-crystallin interaction. A hydrophobic patch mapped between helices 1 and 4 of the FAK FAT domain was found to bind to the β4-β8 groove of αB-crystallin. Such an interaction requires FAK tyrosine 925 and is enhanced following its phosphorylation by Src, which occurs upon FAK stimulation by mechanical stress. αB-crystallin silencing results in calpain-dependent FAK depletion and in increased apoptosis of cardiomyocytes. The overexpression of a myc-FAK construct or treatment with a calpain inhibitor (E64) restored the survival of cardiomyocytes depleted of αB-crystallin. The association between FAK and αB-crystallin was also demonstrated to occur in response to pressure overload in rat left ventricle. The myocardial depletion of αB-crystallin by siRNA results in enhanced apoptosis of cardiomyocytes and myocardial fibrosis in overloaded hearts. These alterations were markedly attenuated in the overloaded left ventricles of transgenic mice with cardiac specific FAK expression. These findings define a novel mechanism by which αB-crystallin controls FAK function, with impact on cardiomyocyte survival and cardiac remodelling in response to stress.

This study was a prospective, parallel-group, randomized, double-blind, vehicle-controlled, multi-centre clinical trial to compare the efficacy of topical sertaconazole 2% cream with vehicle in reducing chronic pruritus in subjects with atopic dermatitis, and to assess its safety and local tolerability. A total of 70 subjects applied either of the 2 treatments twice daily for a period of 4 weeks on affected, itchy skin areas. Treatment efficacy was evaluated primarily considering the item itch intensity on a 5-point verbal rating scale. Insomnia, state of atopic dermatitis (Scoring Atopic Dermatitis; SCORAD), quality of life and therapy benefit were also assessed. No significant difference between active treatment and vehicle was found at any of the time-points for any of the investigated parameters. Under the experimental conditions of the study, sertaconazole 2% cream did not exert anti-pruritic effects that were better than vehicle in subjects with atopic dermatitis who had chronic pruritus. Trial registration ClinicalTrials.gov #NCT01792713.

Ring-shaped bodies are found in the nucleus of Lacandonia schismatica, a rare plant with the sexual organs inverted. They are 0.5-microm-diameter structures that present an electron-dense external ring surrounding a central core. Ultrastructural studies indicate that these bodies contain RNA. The external ring is labeled with antibodies against small nuclear ribonucleoproteins, suggesting that they may be involved in pre-mRNA metabolism. In the present work we further characterized these intranuclear ring-shaped structures by serial-sectioning analysis. Moreover, we tested the presence of additional molecular elements related to pre-mRNA metabolism, such as SR proteins and poly(A)(+) RNA, using immunoelectron microscopy and ultrastructural in situ hybridization. Our results show that these nuclear bodies are spherical. They contain SR proteins involved in splicing and postsplicing events and little to no poly(A)(+) RNA. We also found similar nuclear bodies in other plant and animal species. Therefore, ring-shaped bodies in L. schismatica are spherical, highly compartmentalized nuclear structures that may be involved in pre-mRNA metabolism.

The enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) catalyzes the reutilization of hypoxanthine and guanine to the purine nucleotides IMP and GMP, respectively. HPRT deficiency is an X-linked disorder characterized by uric acid overproduction and variable neurologic impairment. The complete deficiency of HPRT is diagnostic of Lesch-Nyhan syndrome manifested by choreoathetosis, spasticity, mental retardation, and self-injurious behavior. In some HPRT-deficient patients the enzyme defect appeared to be "partial" and the neurologic symptoms mild to severe (Kelley-Seegmiller syndrome). This has prompted the classification of HPRT deficiency in 2 distinct groups: Lesch-Nyhan syndrome and Kelley-Seegmiller syndrome, which has created much confusion. A spectrum of clinical consequences of HPRT deficiency has been recognized in small series of patients, but the complete spectrum of the neurologic disorder has not been described in a single series of patients examined by the same observers. We analyzed our experience with 22 patients belonging to 18 different families with HPRT deficiency diagnosed at "La Paz" University Hospital in Madrid over the past 16 years. The clinical spectrum of these HPRT-deficient Spanish patients was similar to the different phenotypes occasionally reported in the literature, in some cases diagnosed as Lesch-Nyhan "variants." The clinical, biochemical, enzymatic, and molecular genetic studies on these 22 patients allowed us to delineate a new classification of HPRT deficiency. Based on the neurologic symptoms, dependency for personal care, HPRT activity in hemolysate and in intact erythrocytes, and predicted protein size, patients were classified into 4 groups: Group 1 (2 patients), normal development with no neurologic symptoms, HPRT activity was detectable in hemolysates and in intact erythrocytes, and the mutation did not affect the predicted protein size. Group 2 (3 patients) mild neurologic symptoms that did not prevent independent lives, HPRT activity was detectable in intact erythrocytes, and the protein size was normal. Group 3 (2 patients), severe neurologic impairment that precluded an independent life, no residual HPRT activity, and normal protein size. Group 4 (15 patients), clinical characteristics of Lesch-Nyhan syndrome (some may not show self-injurious behavior), no residual HPRT activity, and in most (7 of 8 patients in whom the mutation could be detected) the mutation affected the predicted protein size. This classification of HPRT deficiency into 4 groups may be more useful in terms of accuracy, reproducibility, assessment for treatment trials and prognosis. The study of this Spanish series allows us to conclude that HPRT deficiency may be manifested by a wide spectrum of neurologic symptoms; the overall severity of the disease is associated with mutations permitting some degree of residual enzyme activity; and mutation analysis provides a valuable tool for prognosis, carrier identification, and prenatal diagnosis.

Four computer-typing accommodations were evaluated to determine if arm supports would enhance typing performance and/or comfort of adults with neurological disorders. Each of 12 adults (6 females and 6 males) participated in 7-min typing tasks under the following conditions: 1) without arm support, 2) Ergo Rest arm support, 3) custom arm support in fixed mode, and 4) custom arm support with movable mode. The objective dependent variables were speed, errors, and type of errors. The subjective dependent measures were reported body part discomfort, and arm support ratings. The data were found to be nonparametric in nature. Therefore, nonparametric techniques were employed to analyze the data. Only one of the objective dependent measures were statistically significant; Type of error (additional key). This result may be attributed to the wide range of disability manifestations among participants. Subjective dependent measures were found to be statistically significant, leading to the conclusion that typing with an arm support was more comfortable and easier than without an arm support.

Characteristics of body and fleece weight, and fleece and fibre quality were evaluated in vicunas ( Vicugna vicugna mensalis ) captured for live shearing in Huancavelica, Peru. A total of 305 animals were weighted and shorn between 2003 and 2008. Fibre samples from 104 animals shorn in 2008 were analysed using optical fibre diameter analyser (OFDA) and fibre length measured in a subsample of 68 animals. Mean values for body weight, fleece weight, fibre length and fibre diameter were 37.8 kg, 189.7 g, 30.9 mm and 13.2 μm respectively. Differences due to age were significant for body weight, fleece weight and fibre diameter, but no differences were attributable to sex. The average coefficient of variation of mean fibre diameter was 19.5% showing a higher homogeneity than other luxury animal fibre producers. Mean fibre curvature was medium (79.9°/mm) showing a low relationship with fibre diameter.

Leucine uptake by Saccharomyces cerevisiae is mediated by three transport systems, the general amino acid transport system (GAP), encoded by GAP1, and two group-specific systems (S1 and S2), which also transport isoleucine and valine. A new mutant defective in both group-specific transport activities was isolated by employing a gap1 leu4 strain and selecting for trifluoroleucine-resistant mutants which also showed greatly reduced ability to utilize L-leucine as sole nitrogen source and very low levels of [14C]L-leucine uptake. A multicopy plasmid containing a DNA fragment which complemented the leucine transport defect was isolated by selecting for transformants that grew normally on minimal medium containing leucine as nitrogen source and subsequently assaying [14C]L-leucine uptake. Transformation of one such mutant, lep1, restored sensitivity to trifluoroleucine. The complementing gene, designated LEP1, was subcloned and sequenced. The LEP1 ORF encodes a large protein that lacks characteristics of a transporter or permease (i.e., lacks hydrophobic domains necessary for membrane association). Instead, Lep1p is a very basic protein (pI of 9.2) that contains a putative bipartite signal sequence for targeting to the nucleus, suggesting that it might be a DNA-binding protein. A database search revealed that LEP1 encodes a polypeptide that is identical to Sac3p except for an N-terminal truncation. The original identification of SAC3 was based on the isolation of a mutant allele, sac3-1, that suppresses the temperature-sensitive growth defect of an actin mutant containing the allele act1-1. Sac3p has been previously shown to be localized in the nucleus. When a lep1 mutant was crossed with a sac3 deletion mutant, no complementation was observed, indicating that the two mutations are functionally allelic.

As in many developing nations, basic grains in El Salvador have a critical importance in the rural economy and in the daily diet. Information on price elasticities of supply and demand can be crucial for better informed policy making. This article pre-tests for the suitability of price expectation models and then estimates a market model for each grain. Empirical results point out strong inelasticities in demand and supply for all grains. For maize and red beans (but not for sorghum and rice), low degrees of partial adjustment indicate their high tendency to continue cultivation in spite of adverse economic conditions. © 1998 John Wiley & Sons, Inc.

According to epidemiological studies, gallstonedisease is a very common disease in Mexican-Americansand Mexicans. However, the major risk factors forcholelithiasis in Mexicans have not been identified. We designed a case-control study in a group ofMexican subjects with and without gallstone diseaseconfirmed by ultrasound. These subjects wereprospectively studied over a three-year period. Clinical and epidemiological data were collected bymeans of a questionnaire. A total of 1500 subjects wereincluded in this study: 1000 with and 500 withoutgallstone disease. The major risk factor in both men and women was body mass index [odds ratio (OR)1.64 and 1.96, respectively; P < 0.008 and 0.001]. Inaddition, parity was an important factor in women (OR2.17, P < 0.001), whereas age was associated withgallstone disease in men (OR 1.43, P < 0.001). Wefound that body mass index, parity, and age were theprincipal risk factors for gallstone disease in thisgroup of Mexican subjects. These results are similar tothose found in Mexican-Americanpopulations.

General amino acid permease (GAP1) activity was evaluated in adenylate cyclase-deficient Saccharomyces cerevisiae to determine the effect of cAMP levels on GAP1 activity. Lowering cAMP concentrations in the culture media led to a decrease in the initial rates of L-citrulline uptake. Kinetics of the amino acid transport system showed a partial loss of transport capacity, with no apparent modifications in permease affinity.

We developed a fluorescence in situ hybridization (FISH) method for the rapid detection of Proteus spp. in urine, using a novel peptide nucleic acid (PNA) probe. Testing on 137 urine samples from patients with urinary tract infections has shown specificity and sensitivity values of 98 % (95 % CI, 93.2–99.7) and 100 % (95 % CI, 80,8–100), respectively, when compared with CHROMagar Orientation medium. Results indicate that PNA-FISH is a reliable alternative to traditional culture methods and can reduce the diagnosis time to approximately 2 h., We would like to thank Cristina Silva for providing us the urine samples. This work was supported by Fundacao para a Ciencia e Tecnologia (Postdoctoral Fellowship SFRH/BPD/74480/2010 and project PIC/IC/82815/2007) and by the European Commission's Framework Programme (FP7: "COST-Action TD1004: Theragnostics for imaging and therapy").

Seventy-two long-surviving liver transplant recipients were evaluated prospectively, including a baseline allograft biopsy for weaning off of immunosuppression. Thirteen were removed from candidacy because of chronic rejection (n = 4), hepatitis (n = 2), patient anxiety (n = 5), or lack of cooperation by the local physician (n = 2). The other 59, aged 12-68 years, had stepwise drug weaning with weekly or biweekly monitoring of liver function tests. Their original diagnoses were PBC (n = 9), HCC (n = 1), Wilson's disease (n = 4), hepatitides (n = 15), Laennec's cirrhosis (n = 1), biliary atresia (n = 16), cystic fibrosis (n = 1), hemochromatosis (n = 1), hepatic trauma (n = 1), alpha-1-antitrypsin deficiency (n = 9), and secondary biliary cirrhosis (n = 1). Most of the patients had complications of long-term immunosuppression, of which the most significant were renal dysfunction (n = 8), squamous cell carcinoma (n = 2) or verruca vulgaris of skin (n = 9), osteoporosis and/or arthritis (n = 12), obesity (n = 3), hypertension (n = 11), and opportunistic infections (n = 2). When azathioprine was a third drug, it was stopped first. Otherwise, weaning began with prednisone, using the results of corticotropin stimulation testing as a guide. If adrenal insufficiency was diagnosed, patients reduced to < 5 mg/day prednisone were considered off of steroids. The baseline agents (azathioprine, cyclosporine, or FK506) were then gradually reduced in monthly decrements. Complete weaning was accomplished in 16 patients (27.1%) with 3-19 months drug-free follow-up, is progressing in 28 (47.4%), and failed in 15 (25.4%) without graft losses or demonstrable loss of graft function from the rejections. This and our previous experience with self-weaned and other patients off of immunosuppression indicate that a significant percentage of appropriately selected long-surviving liver recipients can unknowingly achieve drug-free graft acceptance. Such attempts should not be contemplated until 5-10 years posttransplantation and then only with careful case selection, close monitoring, and prompt reinstitution of immunosuppression when necessary.

319 Background: Dose-escalation is a determining factor that influences biochemical control (BC) of prostate cancer. High-dose rate brachytherapy (HDR) is one method for providing BC. Methods: Patients with histological diagnosis Gleason scored (GS), clinical stage T1 to T3a, no evidence of metastatic disease, prostate volume > 20cc < 60cc and initial PSA (PSAi) < 60 mg/ml treated from 1997 to 2005 with an Institutional Policy of treatment were evaluated. Results: There were 273 patients treated with this treatment combination at AC Camargo Cancer Center, Sao Paulo, Brazil. The median age and FU time were 64.7 and 10.3 years, respectively. Two hundred thirteen (78.0%) patients had FU longer than 5 years. Actuarial 10-year OS, CSS and BC were 89.8%, 63.6% and 71.8%, respectively. On univariate analyses GS < 7, clinical stage < T2b, low risk group (LR), absence adjuvant androgen deprivation (ADT), age > 65, PSAi < 10, localized EBRT and three dimensional HDR planning were associated with improved CSS and BC. PSAi and age were not favorable associated to BC. Multivariate Cox regression analyses confirmed LR, GS < 7, PSAi < 10, absence of ADT, age < 65 years as predictors of improved CSS and BC. For OS only LR was confirmed as predictive factor. Conclusions: The present data represents a unique single institution study at long FU for the given technique and the comparison with the current literature data confirms the excellent results achieved with this treatment combination.

A more detailed understanding of the somatic genetic events that drive gastrointestinal adenocarcinomas is necessary to improve diagnosis and therapy. Using data from high-density genomic profiling arrays, we conducted an analysis of somatic copy-number aberrations in 486 gastrointestinal adenocarcinomas including 296 esophageal and gastric cancers. Focal amplifications were substantially more prevalent in gastric/esophageal adenocarcinomas than colorectal tumors. We identified 64 regions of significant recurrent amplification and deletion, some shared and others unique to the adenocarcinoma types examined. Amplified genes were noted in 37% of gastric/esophageal tumors, including in therapeutically targetable kinases such as ERBB2, FGFR1, FGFR2, EGFR, and MET, suggesting the potential use of genomic amplifications as biomarkers to guide therapy of gastric and esophageal cancers where targeted therapeutics have been less developed compared with colorectal cancers. Amplified loci implicated genes with known involvement in carcinogenesis but also pointed to regions harboring potentially novel cancer genes, including a recurrent deletion found in 15% of esophageal tumors where the Runt transcription factor subunit RUNX1 was implicated, including by functional experiments in tissue culture. Together, our results defined genomic features that were common and distinct to various gut-derived adenocarcinomas, potentially informing novel opportunities for targeted therapeutic interventions. Cancer Res; 72(17); 4383–93. ©2012 AACR.

Se presenta una lista de 3,400 especies de plantas vasculares de "La Lacandona", Chiapas, México, que representan el 78.8% del total de especies estimado para la zona. Los datos proceden de la recopilación de identificaciones de material recolectado por el primer autor, de nombres tomados de ejemplares de herbario y de fuentes bibliográficas. Se señalan algunos taxa no registrados anteriormente y se incluyen los sinónimos más comunes. Se presenta una breve delimitación y descripción geográfica de la zona, así como algunos datos de la vegetación y las exploraciones llevadas a cabo en ella.

We studied seven patients aged 14 to 40 years who received living-related kidney transplants and had allograft survivals of 26 to 29 years. The blood urea and creatinine were either within normal limits or marginally elevated. Histopathologic examination showed only mild mesangial expansion, interstitial fibrosis, and arteriosclerosis. Immunoperoxidase staining with anti-HLA antibodies or in situ hybridization with a Y chromosome probe showed persistence of donor tubular epithelium and vascular endothelium within the graft. Recipient-derived glomerular cells were seen in one case, and interstitial lymphocytic infiltrates were seen in all cases. A review of the clinicopathologic data available for these cases indicated that both central and peripheral immunologic mechanisms contributed to the maintenance of prolonged graft survival. This extended survival was independent of six antigen matching, down-regulation of donor HLA antigen expression, and ingrowth of host epithelium/endothelium into the allograft.

The effect of ergosterol depletion by ketoconazole on the leishmanicidal activity of the pore-forming antibiotic amphotericin B (AmB) was investigated. Leishmania mexicana promastigotes were lysed within minutes by the addition of micromolar concentrations of AmB (0.5 microM) but became insensitive to AmB after growth in the presence of ketoconazole (0.25 microM, 90 h). Lipid chromatographic analysis indicated that under such conditions, ketoconazole depleted the major Leishmania sterols, dehydroepisterol and ergosterol. Plasma membrane vesicles prepared from ketoconazole-treated promastigotes exhibited a much reduced enhancement of their salt permeability after the addition of AmB at concentrations as high as 5 microM. This finding clearly indicates that upon ketoconazole treatment, the capacity of pore formation by the antibiotic is substantially impaired. The reduction of desmethyl sterols by ketoconazole was accompanied by a significant increase of 14-alpha-methyl sterols, but exogenous cholesterol remained unchanged. This ability of Leishmania promastigotes to incorporate cholesterol from the external medium may explain why ketoconazole-treated cells exhibited a much decreased but significative response to AmB when they were exposed to high AmB concentrations (2.5 or 5.0 microM). Parallel measurements by using a fluorescence energy transfer method indicated that binding of AmB to ketoconazole-treated Leishmania promastigotes and heat-transformed leishmanias was also decreased but to different extents, a finding that may be related to the differences in their sterol content. The results obtained clearly indicate that the specific interaction of AmB with desmethyl sterols, such as dehydroepisterol, ergosterol, and even exogenous cholesterol, is an absolute requirement for the lethal action exerted by this polyene antibiotic on L. mexicana promastigotes.

Summary The interphase nucleus of the cells of several tissues of Lacandonia schismatica was studied using electron microscopy cytochemical and immunocytochemical methods. The EDTA staining procedure, preferential for RNP, contrasted the Lacandonia granules and perichromatin fibrils. These granules were found to be relatively resistant to RNAse hydrolysis, but they were easily digested if RNAse treatment was carried out after a short hydrolysis with pronase. Bismuth oxynitrate stained granular structures about 17 nm in diameter and the periphery of a few Lacandonia granules. The anti=snURNPs bound to RNP-containing fibrils in the perichromatin and interchromatin space and also to the periphery of some Lacandonia granules. Immunolabeling of DNA demonstrated numerous filaments of extended chromatin in the perichromatin and interchromatin spaces which were closely related to Lacandonia granules. These observations suggested that Lacandonia granules are equivalent to Balbiani ring granules of nuclei with polytene chromosomes and to perichromatin granules of other plant and animal nuclei. The small number of Lacandonia granules labeled in their periphery by anti-snURNP mAb were interpreted as being immature granules in the process of formation. The external or annular part of the ring-shaped structures is heavily labeled by anti-URNP mAbs but scarcely stained by the EDTA procedure. These features indicate that this region contains abundant proteins associated with snURNAs but probably little snURNAs. The synaptonemal-like complexes previously found in the interphase nuclei of Lacandonia are formed by two parallel masses of compact chromatin, which react with anti-DNA, and a central clear space crossed by fiber. Most of these fibers do not react with tests for DNA or RNA and are probably composed of proteins. The EDTA procedure contrasts some fibrils at the internal aspect of the masses of compact chromatin, suggesting the presence of remnants of the lateral element of a synaptonemal complex.

A case of giant condyloma acuminatum of the scrotum is reported. It was initially diagnosed as a low grade squamous carcinoma. This neoplasm is closely related to Human Papillomavirus, mainly HPV 6 and HPV 11, and may be clinically indistinguishable from squamous carcinoma. The histological appearance is benign, however, its clinical behaviour is similar to verrucous carcinoma. A review of previously reported cases has been undertaken. The true significance of this lesion and management are discussed. Early aggressive therapy is recommended.

A prospective randomized, double-blind, double-dummy study investigated the effects of two angiotensin converting enzyme (ACE) inhibitors on urinary albumin excretion in nondiabetic patients with mild to moderate essential hypertension. At the end of a 4-week placebo run-in period, 36 patients were randomly allocated to receive a 12-week course of treatment with quinapril (10, 20, or 40 mg) once daily or captopril (25, 50, or 75 mg) twice daily. Seventeen patients in each group completed the study. The mean change in mean blood pressure for patients taking quinapril (mean dose 32 mg/24 h) was -5 mm Hg (P = .002), and for patients taking captopril (mean dose 132 mg/24 h), -9 mm Hg (P = .002). The baseline urinary albumin excretion rate in both groups was (mean +/- EEM) 57 +/- 7 micrograms/min. Fifteen patients in the quinapril group and 12 patients in the captopril group had baseline albumin excretion rates of more than 20 micrograms/min. Mean urinary albumin excretion decreased in patients treated with quinapril from 55 to 33 micrograms/min (mean decrease 22 micrograms/min, 95% confidence interval [CI], 1 to 43 micrograms/min; P = .031) and with captopril from 59 to 41 micrograms/min (mean decrease 18 micrograms/min, 95% CI, 3 to 32 micrograms/min; P = .025). No significant modifications were observed in serum lipid concentrations, serum and urinary electrolytes, and magnesium or phosphorus metabolism. Mean urinary calcium excretion decreased in the quinapril-treated group (from 219 to 188 mg/24 h; P = .023) but not in the captopril group (from 264 to 267 mg/24 h).(ABSTRACT TRUNCATED AT 250 WORDS)

Prostate cancer is the second most common cancer in men worldwide and causes over 250,000 deaths each year. Overtreatment of indolent disease also results in significant morbidity. Common genetic alterations in prostate cancer include losses of NKX3.1 (8p21) and PTEN (10q23), gains of AR (the androgen receptor gene) and fusion of ETS family transcription factor genes with androgen-responsive promoters. Recurrent somatic base-pair substitutions are believed to be less contributory in prostate tumorigenesis but have not been systematically analyzed in large cohorts. Here, we sequenced the exomes of 112 prostate tumor and normal tissue pairs. New recurrent mutations were identified in multiple genes, including MED12 and FOXA1. SPOP was the most frequently mutated gene, with mutations involving the SPOP substrate-binding cleft in 6–15% of tumors across multiple independent cohorts. Prostate cancers with mutant SPOP lacked ETS family gene rearrangements and showed a distinct pattern of genomic alterations. Thus, SPOP mutations may define a new molecular subtype of prostate cancer., National Human Genome Research Institute (U.S.) (Large Scale Sequencing Program Grant U54 HG003067)

To characterize somatic alterations in colorectal carcinoma, we conducted a genome-scale analysis of 276 samples, analysing exome sequence, DNA copy number, promoter methylation and messenger RNA and microRNA expression. A subset of these samples (97) underwent low-depth-of-coverage whole-genome sequencing. In total, 16% of colorectal carcinomas were found to be hypermutated: three-quarters of these had the expected high microsatellite instability, usually with hypermethylation and MLH1 silencing, and one-quarter had somatic mismatch-repair gene and polymerase ε (POLE) mutations. Excluding the hypermutated cancers, colon and rectum cancers were found to have considerably similar patterns of genomic alteration. Twenty-four genes were significantly mutated, and in addition to the expected APC, TP53, SMAD4, PIK3CA and KRAS mutations, we found frequent mutations in ARID1A, SOX9 and FAM123B. Recurrent copy-number alterations include potentially drug-targetable amplifications of ERBB2 and newly discovered amplification of IGF2. Recurrent chromosomal translocations include the fusion of NAV2 and WNT pathway member TCF7L1. Integrative analyses suggest new markers for aggressive colorectal carcinoma and an important role for MYC-directed transcriptional activation and repression., National Institutes of Health (U.S.) (Grant U24CA143799), National Institutes of Health (U.S.) (Grant U24CA143835), National Institutes of Health (U.S.) (Grant U24CA143840), National Institutes of Health (U.S.) (Grant U24CA143843), National Institutes of Health (U.S.) (Grant U24CA143845), National Institutes of Health (U.S.) (Grant U24CA143848), National Institutes of Health (U.S.) (Grant U24CA143858), National Institutes of Health (U.S.) (Grant U24CA143866), National Institutes of Health (U.S.) (Grant U24CA143867), National Institutes of Health (U.S.) (Grant U24CA143882), National Institutes of Health (U.S.) (Grant U24CA143883), National Institutes of Health (U.S.) (Grant U24CA144025), National Institutes of Health (U.S.) (Grant U54HG003067), National Institutes of Health (U.S.) (Grant U54HG003079), National Institutes of Health (U.S.) (Grant U54HG003273)

Continuous culture and in vivo experiments were conducted to measure changes in ruminal fermentation and animal performance when crude glycerol was added to diets. For the continuous culture experiment (n = 6), diets consisted of 4 levels of crude glycerol (0, 5, 10, and 20%) that replaced corn grain. Dry matter and OM digestibility decreased linearly (P0.05) when crude glycerol increased in the diet, and no effect (P = 0.20 and 0.65, respectively) was observed for CP and NDF digestibility. Total VFA concentration and ammonia did not change (P0.05) due to crude glycerol level. Microbial efficiency increased quadratically (P = 0.012) as crude glycerol increased, whereas microbial N flow did not differ (P = 0.36) among treatments. As crude glycerol increased in the diet, crude glycerol digestibility decreased (P0.05). Seventy-two crossbred steer calves (250 ± 2.0 kg) were assigned to 4 treatments: 0, 5, 10, and 20% crude glycerol that replaced corn grain. Animals were fed for a total of 150 d. No differences (P = 0.08) between treatments were measured for DMI. Average daily gain and GF responded quadratically (P0.05), with 10% crude glycerol resulting in the greatest values. In the second in vivo experiment, 100 crossbred steer calves (300 ± 2.0 kg) were assigned to 5 treatments: 0, 5, 10, 12.5, or 15% crude glycerol replaced corn grain. Calves were fed for a total of 135 d. No significant differences (P0.05) were measured in growth performance. For Exp. 3, one hundred heifer calves (270 ± 2.0 kg) were assigned to 4 treatments: 0, 5, 10, or 20% crude glycerol that replaced hay. No differences (P0.05) were measured in animal performance. We concluded that crude glycerol addition to a diet did not negatively affect ruminal fermentation, and addition of up to 20% in concentrate and hay-based diets should not affect performance or carcass characteristics.

2012 August 09 Author Manuscript, Head and neck squamous cell carcinoma (HNSCC) is a common, morbid, and frequently lethal malignancy. To uncover its mutational spectrum, we analyzed whole-exome sequencing data from 74 tumor-normal pairs. The majority exhibited a mutational profile consistent with tobacco exposure; human papillomavirus was detectable by sequencing DNA from infected tumors. In addition to identifying previously known HNSCC genes (TP53, CDKN2A, PTEN, PIK3CA, and HRAS), our analysis revealed many genes not previously implicated in this malignancy. At least 30% of cases harbored mutations in genes that regulate squamous differentiation (for example, NOTCH1, IRF6, and TP63), implicating its dysregulation as a major driver of HNSCC carcinogenesis. More generally, the results indicate the ability of large-scale sequencing to reveal fundamental tumorigenic mechanisms.

High concentrations of long-chain fatty acids have been found to be harmful to mammalian cells and prokaryotic organisms. This effect was investigated in Saccharomyces cerevisiae. Addition of 3 mmol/L palmitate to a yeast extract-peptone medium caused a significant inhibition of cell growth during the first 2 d of incubation, followed by renewed growth and palmitate utilization. Inhibition was also observed with palmitate concentrations down to 0.1 mmol/L. As inferred from catalase activity determinations, this effect was found to correlate with the absence of peroxisome proliferation. Finally, no inhibition was observed in exponential-phase cultures or in the presence of 0.1 g/L glucose, this suggesting that the physiological state of the cell may determine whether its growth will be inhibited by fatty acids.

Background Squamous cell lung carcinomas account for approximately 25% of new lung carcinoma cases and 40,000 deaths per year in the United States. Although there are multiple genomically targeted therapies for lung adenocarcinoma, none has yet been reported in squamous cell lung carcinoma. Methodology/Principal Findings Using SNP array analysis, we found that a region of chromosome segment 8p11-12 containing three genes–WHSC1L1, LETM2, and FGFR1–is amplified in 3% of lung adenocarcinomas and 21% of squamous cell lung carcinomas. Furthermore, we demonstrated that a non-small cell lung carcinoma cell line harboring focal amplification of FGFR1 is dependent on FGFR1 activity for cell growth, as treatment of this cell line either with FGFR1-specific shRNAs or with FGFR small molecule enzymatic inhibitors leads to cell growth inhibition. Conclusions/Significance These studies show that FGFR1 amplification is common in squamous cell lung cancer, and that FGFR1 may represent a promising therapeutic target in non-small cell lung cancer., Novartis Pharmaceuticals Corporation, American Lung Association, Uniting Against Lung Cancer, Sara Thomas Monopoli Fund, Seaman Foundation, India. Dept. of Biotechnology, National Lung Cancer Partnership

Background & Aim. Non alcoholic fatty liver disease (NAFLD) is the most common liver disease in Western countries. Population studies have demonstrated that men and posmenopausal women have higher prevalence of NAFLD. The aim was to investigate the prevalence of NAFLD in premenopausal, posmenopausal and polycystic ovary syndrome (PCOS) women. Methods. A cross sectional study carried out at University Hospital in Mexico City from January 2009 to November 2009. One hundred ninety seven women who agreed to participate were divided into groups, comprising 93 with NAFLD and without NAFLD. Anthropometric, metabolic and biochemical variables were measured. Serum estradiol and cortisol concentrations were determined and compared between the groups. Results. Of the 197 patients, 93(47.2%) had NAFLD and 104 (52.8%) did not have NAFLD. The prevalence of NAFLD in premenopausal, postmenopausal and PCOS patients was 32.2, 57.9, and 62%, respectively. Age, BMI, hip to waist ratio, fasting glucose, HOMA –IR, and insulin were significantly higher in NAFLD patients. Women without NAFLD had significantly higher levels of serum estradiol (100 ± 95.4) compared with NAFLD patients (55.5 ± 66.6) p = 0.001. By group with and without NAFLD: premenopausal (55.44±93.3 vs. 128.56 ± 109.22), posmenopausal (44.98 ± 51.41 vs. 42.72 ± 51.48) and PCOS women (64.9 ± 53.3 vs. 101.36 ± 80.89) had significantly different hormone profile. Conclusion. These results suggest that NAFLD is more prevalent in postmenopausal and women with PCOS than those premenopausal ones. The estrogens may have a protective effect of against NAFLD in women.

Melanoma is notable for its metastatic propensity, lethality in the advanced setting and association with ultraviolet exposure early in life. To obtain a comprehensive genomic view of melanoma in humans, we sequenced the genomes of 25 metastatic melanomas and matched germline DNA. A wide range of point mutation rates was observed: lowest in melanomas whose primaries arose on non-ultraviolet-exposed hairless skin of the extremities (3 and 14 per megabase (Mb) of genome), intermediate in those originating from hair-bearing skin of the trunk (5–55 per Mb), and highest in a patient with a documented history of chronic sun exposure (111 per Mb). Analysis of whole-genome sequence data identified PREX2 (phosphatidylinositol-3,4,5-trisphosphate-dependent Rac exchange factor 2)—a PTEN-interacting protein and negative regulator of PTEN in breast cancer—as a significantly mutated gene with a mutation frequency of approximately 14% in an independent extension cohort of 107 human melanomas. PREX2 mutations are biologically relevant, as ectopic expression of mutant PREX2 accelerated tumour formation of immortalized human melanocytes in vivo. Thus, whole-genome sequencing of human melanoma tumours revealed genomic evidence of ultraviolet pathogenesis and discovered a new recurrently mutated gene in melanoma., National Human Genome Research Institute (U.S.)

Multiple myeloma is an incurable malignancy of plasma cells, and its pathogenesis is poorly understood. Here we report the massively parallel sequencing of 38 tumour genomes and their comparison to matched normal DNAs. Several new and unexpected oncogenic mechanisms were suggested by the pattern of somatic mutation across the data set. These include the mutation of genes involved in protein translation (seen in nearly half of the patients), genes involved in histone methylation, and genes involved in blood coagulation. In addition, a broader than anticipated role of NF-κB signalling was indicated by mutations in 11 members of the NF-κB pathway. Of potential immediate clinical relevance, activating mutations of the kinase BRAF were observed in 4% of patients, suggesting the evaluation of BRAF inhibitors in multiple myeloma clinical trials. These results indicate that cancer genome sequencing of large collections of samples will yield new insights into cancer not anticipated by existing knowledge., Multiple Myeloma Research Foundation

2011 August 10., Prostate cancer is the second most common cause of male cancer deaths in the United States. However, the full range of prostate cancer genomic alterations is incompletely characterized. Here we present the complete sequence of seven primary human prostate cancers and their paired normal counterparts. Several tumours contained complex chains of balanced (that is, ‘copy-neutral’) rearrangements that occurred within or adjacent to known cancer genes. Rearrangement breakpoints were enriched near open chromatin, androgen receptor and ERG DNA binding sites in the setting of the ETS gene fusion TMPRSS2–ERG, but inversely correlated with these regions in tumours lacking ETS fusions. This observation suggests a link between chromatin or transcriptional regulation and the genesis of genomic aberrations. Three tumours contained rearrangements that disrupted CADM2, and four harboured events disrupting either PTEN (unbalanced events), a prostate tumour suppressor, or MAGI2 (balanced events), a PTEN interacting protein not previously implicated in prostate tumorigenesis. Thus, genomic rearrangements may arise from transcriptional or chromatin aberrancies and engage prostate tumorigenic mechanisms., National Human Genome Research Institute (U.S.)

The use of preparations from Bryophyllum pinnatum in tocolysis is supported by both clinical (retrospective comparative studies) and experimental (using uterus strips) evidence. We studied here the effect of B. pinnatum juice on the response of cultured human myometrial cells to stimulation by oxytocin, a hormone known to be involved in the control of uterine contractions by increasing the intracellular free calcium concentration ([Ca2+]i). In this work, [Ca2+]i was measured online during stimulation of human myometrial cells (hTERT-C3 and M11) with oxytocin, which had been pre-incubated in the absence or in the presence of B. pinnatum juice. Since no functional voltage-gated Ca2+ channels could be detected in these myometrial cells, the effect of B. pinnatum juice was as well studied in SH-SY5Y neuroblastoma cells, which are known to have such channels and can be depolarised with KCl. B. pinnatum juice prevented the oxytocin-induced increase in [Ca2+]i in hTERT-C3 human myometrial cells in a dose-dependent manner, achieving a ca. 80% inhibition at a 2% concentration. Comparable results were obtained with M11 human primary myometrial cells. In hTERT-C3 cells, prevention of the oxytocin-induced increase in [Ca2+]i was independent of the extracellular Ca2+ concentration and of voltage-dependent Ca2+-channels. B. pinnatum juice delayed, but did not prevent the depolarization-induced increase in [Ca2+]i in SH-SY5Y cells. Taken together, the data suggest a specific and concentration-dependent effect of B. pinnatum juice on the oxytocin signalling pathway, which seems to corroborate its use in tocolysis. Such a specific mechanism would explain the rare and minor side-effects in tocolysis with B. pinnatum as well as its high therapeutic index.

Background It is well established that adaptive immune responses induced by hypercholesterolemia play an important role in the development of atherosclerosis, but the pathways involved remain to be fully characterized. In the present study we assessed immune responses to hypercholesterolemia induced by feeding Apoe-/- mice a high-fat diet for 4 or 8 weeks. Results The primary immune response in lymph nodes draining the aortic root was an increased expression of interferon (IFN)-γ in CD8+CD28+ T cells, while an activation of IFN-γ expression in CD4+ T cells was observed only after 8 weeks of high-fat diet. Contrarily, spleen CD4+ T cells responded with a higher expression of IL-10. Spleen CD8+ T cells expressed both IFN-γ and IL-10 and showed enhanced proliferation when exposed to Concanavalin A. Plasma levels of IgG and IgM against oxidized LDL did not change, but the level of apolipoprotein B/IgM immune complexes was increased. Conclusion Hypercholesterolemia leads to unopposed activation of Th1 immune responses in lymph nodes draining atherosclerotic lesions, whereas Th1 activation in the spleen is balanced by a concomitant activation of Th2 cells. The activation of CD8+ T cells implies that hypercholesterolemia is associated with formation of cell autoantigens.

By light and electron microscope cytochemistry we characterized the interphase nucleus ofLacandonia schismatica, the only known species of the new plant family Lacandoniaceae, whose most peculiar feature is the inverted position of the sexual organs, an aspect never found before among flowering plants. Furthermore, we compare it toTriuris alata, a related species, toVoyria aphylla (a dicotyledon), toGymnosiphon divaricatus (a monocotyledon) and also to saprophytes. The reticulated chromatin ofL schismatica andT alata is similar to that of other monocotyledons. In addition, we describe a unique type of RNP granules in the interchromatin space which are about 32 ± 3 nm SD in diameter and occur as huge clusters. They are intermediate in size and spatial distribution between inter- and peri-chromatin granules. We term them ‘Lacandonia granules’. The granules were also found inT alata. They are 31 ± 2 nm in diameter. No significant differences in size were observed between them (P > 0.05). Synaptonemallike complexes and ring-shaped structures were seen in interphase nuclei of somatic cells of these species. Coiled and nucleolus-associated bodies, as well as centromeres were also found in these two organisms. On the contrary,Vaphylla andG divaricatus display a chromocentric nuclear organization. The nuclear similarities betweenL schismatica andT alata suggest extremely close phylogenetic relationships between them.

As all hydrological models are intrinsically limited hypotheses on the behaviour of catchments, models – which attempt to represent real-world behaviours – will always remain perfectible. To progress on the long road towards improved models, we need demanding tests, i.e. true crash tests. Efficient development and assessment of hydrological models requires demanding tests on large and varied data sets to ensure models' generality, to diagnose their failures and limitations, and ultimately, to improve them.

Five patients with mild chronic portal systemic encephalopathy (PSE) were studied. The study was designed in a double cross over fashion in which each patient received during period I a 40 g vegetable protein diet as single treatment. During period II three g/day of oral kanamycin were added and then new periods of single vegetable protein diet (period III) and vegetable protein diet plus kanamycin (period IV) were introduced (identical to periods I and II respectively). Each period lasted two weeks. Several biweekly assessements-tests were determined including: mental state, asterixis grade, electroencephalograms, number connection tests, figure connection tests, blood ammonia levels and stool counts of total aerobes/anaerobes per g/feces were done. During the study none of the patients developed acute encephalopathy. In any case it was detected a significant improvement of the PSE parameters assessed with the addition of oral kanamycin. Fecal counts were very similar during the various periods of the study. We conclude that in mild portal systemic encephalopathy controlled with vegetable protein diet, the addition of non absorbable antibiotics is not mandatory for the management of these patients and may represent a potential risk of serious side effects. At the beginning of treatment vegetable protein diet should be administered and only in case of failure, antibiotics are to be indicated.

Lineage survival oncogenes are activated by somatic DNA alterations in cancers arising from the cell lineages in which these genes play a role in normal development.1,2 Here we show that a peak of genomic amplification on chromosome 3q26.33, found in squamous cell carcinomas (SCCs) of the lung and esophagus, contains the transcription factor gene SOX2—which is mutated in hereditary human esophageal malformations3 and necessary for normal esophageal squamous development4, promotes differentiation and proliferation of basal tracheal cells5 and co-operates in induction of pluripotent stem cells.6,7,8 SOX2 expression is required for proliferation and anchorage-independent growth of lung and esophageal cell lines, as shown by RNA interference experiments. Furthermore, ectopic expression of SOX2 cooperated with FOXE1 or FGFR2 to transform immortalized tracheobronchial epithelial cells. SOX2-driven tumors show expression of markers of both squamous differentiation and pluripotency. These observations identify SOX2 as a novel lineage survival oncogene in lung and esophageal SCC.

New chiral amino-alcohols were enantioselectively synthesized using biotransformations as the key steps. They were used as ligand in the enantioselective borane reduction of acetophenone and of the corresponding anti oxime methyl ether.

Objective: To present our experience with the anterolateral thigh flap in reconstructing a full thickness defect of the buccal mucosa and cheek. Methods: Design: Case Report Setting: Tertiary Private Hospital Patient: One Results: A 36 year old male with a T4aN0M0 Stage IVa buccal carcinoma on the left underwent wide excision, marginal mandibulectomy and modified radical neck dissection with preservation of the spinal accessory nerve, internal jugular vein and sternocleidomastoid muscle. The resultant through-and-through defect of the cheek skin measuring 8 x 6cm and buccal mucosa measuring 6 x 10 cm with a concomitant ¼ upper lip and 1/3 lower lip defect was reconstructed with an anterolateral thigh free flap. The patient recovered uneventfully and underwent adjuvant concurrent chemoradiotherapy 1 month post-operation. At 2 ½ months post-operation, he had no oral incontinence and could resume a normal diet with good speech. Conclusion: The anterolateral thigh free flap is an excellent soft-tissue flap for reconstruction of a full thickness defect of the buccal area. Key words: Surgical flap, Carcinoma, squamous cell, Oral cancer

Objective: To determine the antimycotic activity of the four medicinal plant extracts, kalachuchi bark (Plumeria acuminata Ait.), atsuete bark (Bixa orellana Linn.), akapulko leaves (Cassia alata Linn.), and neem leaves (Azadirachta indica Adr. Juss), when compared to the standard clotrimazole in the treatment of otomycosis. Study Design: Experimental Study Methods: Taxonomically identified plants, kalachuchi, atsuete, akapulko, and neem tree were collected and deposited in an herbarium. Extracts of these plants and the standard clotrimazole were tested against isolates of Aspergillus flavus, Aspergillus niger, and Candida albicans taken from patients with otomycosis. Three trials were made for each extract using different solvents and results subjected to statistical analysis. Result: Of the four medicinal plant extracts studied, only kalachuchi bark extract exhibited antifungal activity against Aspergillus flavus and Aspergillus niger using methylethylketone as solvent when compared to the standard clotrimazole. It was equally effective in inhibiting the growth of A. flavus and A. niger. However, all plant extracts using all types of solvents were equally ineffective in inhibiting the growth of Candida albicans. Conclusion: This in vitro study suggested that kalachuchi (Plumeria acuminata Linn.) bark extract inhibits the growth of Aspergillus species and was comparable to the standard clotrimazole. Following appropriate further studies and clinical trials, it may be a potential alternative treatment option for otomycosis caused by Aspergillus species. Key words: otomycosis; kalachuchi (Plumeria acuminata Linn.) bark; antimycotic; Aspergillus flavus; Aspergillus niger

A comparative study of the effect of the polyene antibiotic amphotericin B (AmB) on the viability of Leishmania mexicana promastigotes before and after their transformation by heat into amastigotelike forms was carried out. The kinetics of cell death were followed by spectrofluorometry with the nucleic acid-binding compound ethidium bromide. It was found that the rapid killing effect that is exerted by AmB on Leishmania promastigotes was even faster after their transformation into amastigotelike forms. Binding studies of AmB to Leishmania membranes by circular dichroism indicated that heat transformation modified it from noncooperative to cooperative binding, decreasing the amount of antibiotic that bound to the membranes. Thus, the increased rate of ethidium bromide incorporation into transformed cells was not related either to the amount of AmB bound or to an increased amount of ergosterol in the membrane (the ergosterol/phospholipid ratio was four times smaller after heat shock). An increase in the Mg2+ content of the external aqueous solution was able to prevent the AmB-induced incorporation of ethidium bromide into Leishmania promastigotes to a greater extent (Ki = 13.8 mM) than it was into heat-transformed cells (Ki = 64 mM), suggesting that there were significant changes at the Leishmania cell surface on heat transformation. The significance of these results for understanding the mechanism of action of AmB on sensitive organisms is discussed.

Gallstone disease (GD) and cardiovascular disease (CD) are common diseases worldwide with considerable economical impact and they are strongly associated. Carotid atherosclerosis is an excellent marker of risk for CD like stroke and myocardial infarction. The aim of this study was to assess the association between gallstones and carotid atherosclerosis.A cross-sectional study was conducted. We evaluated subjects with ultrasonographical evidence of GD and asymptomatic subjects without such evidence. Anthropometric, clinical and biochemical variables were collected. The Metabolic syndrome was evaluated using adult treatment panel III criteria. Carotid artery intima-media thickness (CIMT) was determined by a standard ultrasound protocol. Insulin-like growth factor-1 (IGF-1) serum levels were measured in all subjects.We studied 191 subjects: 62 subjects with GD (53.2% males) and 129 asymptomatic subjects without GD (65.9% males). Subjects with GD exhibited a higher body mass index, body fat percent, insulin serum levels and CIMT (P0.05 for all). The prevalence of GD was higher in subjects with a CIMT0.75 independently of other factors [odds ratio (OR) 2.12, 95% confidence interval (CI) 1.04-4.34; P=0.039], and for every 0.1 mm increase in CIMT the independent probability to be a case of GD increased by a factor of 1.25 (95% CI 1.02-1.53; P=0.027). IGF-1 levels did not differ among groups.Subjects with GD exhibit greater carotid atherosclerosis, and therefore have a higher risk for stroke and myocardial infarction.