Casein kinase 2 phosphorylates and induces the SALL2 tumor suppressor degradation in colon cancer cells.

Spalt-like proteins are Zinc finger transcription factors from Caenorhabditis elegans to vertebrates, with critical roles in development. In vertebrates, four paralogues have been identified (SALL1-4), and SALL2 is the family's most dissimilar member. SALL2 is required during brain and eye development. It is downregulated in cancer and acts as a tumor suppressor, promoting cell cycle arrest and cell death. Despite its critical functions, information about SALL2 regulation is scarce. Public data indicate that SALL2 is ubiquitinated and phosphorylated in several residues along the protein, but the mechanisms, biological consequences, and enzymes responsible for these modifications remain unknown. Bioinformatic analyses identified several putative phosphorylation sites for Casein Kinase II (CK2) located within a highly conserved C-terminal PEST degradation motif of SALL2. CK2 is a serine/threonine kinase that promotes cell proliferation and survival and is often hyperactivated in cancer. We demonstrated that CK2 phosphorylates SALL2 residues S763, T778, S802, and S806 and promotes SALL2 degradation by the proteasome. Accordingly, pharmacological inhibition of CK2 with Silmitasertib (CX-4945) restored endogenous SALL2 protein levels in SALL2-deficient breast MDA-MB-231, lung H1299, and colon SW480 cancer cells. Silmitasertib induced a methuosis-like phenotype and cell death in SW480 cells. However, the phenotype was significantly attenuated in CRISPr/Cas9-mediated SALL2 knockout SW480 cells. Similarly, Sall2-deficient tumor organoids were more resistant to Silmitasertib-induced cell death, confirming that SALL2 sensitizes cancer cells to CK2 inhibition. We identified a novel CK2-dependent mechanism for SALL2 regulation and provided new insights into the interplay between these two proteins and their role in cell survival and proliferation.

Mating-based regulation and ligand binding of an odorant-binding protein support the inverse sexual communication of the greater wax moth, Galleria mellonella (Lepidoptera: Pyralidae).

In moths, sex pheromones play a key role in mate finding. These chemicals are transported in the antennae by odorant-binding proteins (OBPs). Commonly, males encounter conspecific females; therefore, several OBPs are male-biased. Less is known, however, about how the olfactory system of moths has evolved toward inverse sexual communication, ie where females seek males. Therefore, the objective of this study was to identify the profile of OBPs and their expression patterns in the bee hive pest, Galleria mellonella, a moth that uses inverse sexual communication. Here, OBP-related transcripts were identified by an RNA Sequencing (RNA-Seq) approach and analysed through both Reverse Transcription Polymerase Chain Reaction (RT-PCR) in different tissues and quantitative real-time PCR for two states, virgin and postmating. Our results indicate that G. mellonella has 20 OBPs distributed amongst different tissues. Interestingly, 17 of the 20 OBPs were significantly down-regulated after mating in females, whereas only OBP7 was up-regulated. By contrast, 18 OBP transcripts were up-regulated in males after mating. Additionally, binding assays and structural simulations showed general odorant-binding protein 2 (GOBP2) was able to bind sex pheromone components and analogues. These findings suggest a possible role of OBPs, especially GOBPs, in the inverse sexual communication of G. mellonella, with gene expression regulated as a response to mating.

Raman Spectroscopy and Chemometric Modeling to Predict Physical-Chemical Honey Properties from Campeche, Mexico.

In this work, 10 chemometric models based on Raman spectroscopy were constructed to predict the physicochemical properties of honey produced in the state of Campeche, Mexico. The properties of honey studied were pH, moisture, total soluble solids (TSS), free acidity, lactonic acidity, total acidity, electrical conductivity, Redox potential, hydroxymethylfurfural (HMF), and ash content. These proprieties were obtained according to the methods described by the Association of Official Analytical Chemists, Codex Alimentarius, and the International Honey Commission. For the construction of the chemometric models, 189 honey samples were collected and analyzed in triplicate using Raman spectroscopy to generate the matrix data [X], which were correlated with each of the physicochemical properties [Y]. The predictive capacity of each model was determined by cross validation and external validation, using the statistical parameters: standard error of calibration (SEC), standard error of prediction (SEP), coefficient of determination of cross-validation (R2cal), coefficient of determination for external validation (R2val), and Student's t-test. The statistical results indicated that the chemometric models satisfactorily predict the humidity, TSS, free acidity, lactonic acidity, total acidity, and Redox potential. However, the models for electric conductivity and pH presented an acceptable prediction capacity but not adequate to supply the conventional processes, while the models for predicting ash content and HMF were not satisfactory. The developed models represent a low-cost tool to analyze the quality of honey, and contribute significantly to increasing the honey distribution and subsequently the economy of the region.

Antennal Morphology and Localization of a Pheromone-Binding Protein of Lobesia botrana (Denis & Schiffermüller) (Lepidoptera: Tortricidae).

In the sensory system of insects, olfactory sensilla constitute important functional elements for discriminating odors. Therefore, we used light microscopy and scanning electron microscopy to investigate the morphology and distribution of sensilla in the antennae of Lobesia botrana (Denis & Schiffermüller). In addition, we studied the expression of the gene encoding for pheromone-binding protein 1 (LbotPBP1) by in situ hybridization. Lobesia botrana antennae are filiform and are subdivided into three segments: scape, pedicel, and flagellum. The number of flagellum and their overall length were significantly higher and longer in males than in females. Six morphological types of sensilla (trichodea, chaetica, coeloconica, auricillica, basiconica, and styloconica) were identified on the antennae of both sexes. Trichodea sensilla were the most abundant on the antennae of L. botrana, and three subtypes, discerned by their lengths, were observed. However, sensilla trichodea subtype III was only present in male antennae. Moreover, LbotPBP1 expression was restricted to this type of sensilla, thus confirming its olfactory role, specifically under the context of sexual pheromone perception.

The Effect of Protease Inhibitors on Digestive Proteolytic Activity in the Raspberry Weevil, Aegorhinus superciliosus (Guérin) (Coleoptera: Curculionidae).

The raspberry weevil, Aegorhinus superciliosus (Guérin) (Coleoptera: Curculionidae), is an economically important pest of blueberry in southern Chile. The digestive protease activity of adult insects was investigated using general and specific substrates and inhibitors. Enzymatic assays demonstrated the presence of trypsin- and chymotrypsin-like serine proteinases. Furthermore, in vitro assays using phenylmethylsulfonyl fluoride (PMSF) and soybean trypsin inhibitor (SBTI) at 0.01 and 0.1 mM showed percentages of enzymatic inhibition between 0 and 16% for PMSF and 67 to 76% for SBTI, whereas in vivo assays indicated that SBTI caused between 50 and 90% mortality in males and between 80 and 100% in females. Our data indicate the presence of serine proteases and suggest that digestive proteases could be a target for the design and development of strategies to control the raspberry weevil.

Repellent Effect and Metabolite Volatile Profile of the Essential Oil of Achillea millefolium Against Aegorhinus nodipennis (Hope) (Coleoptera: Curculionidae).

Aegorhinus nodipennis (Hope) (Coleoptera: Curculionidae) is an important native pest in fruit crops that is mainly found in European hazelnut fields in the south of Chile. We investigated the behavioral response of A. nodipennis to volatile compounds released from the essential oil of Achillea millefolium and its main constituent using olfactometric bioassays. Gas chromatographic and mass spectral analysis of the A. millefolium essential oil revealed the presence of 11 compounds. Monoterpene ß-thujone (96.2%) was the main component of the oil. Other compounds identified were α-thujone, 1,8-cineole, p-cymene, and 4-terpineol, all with percentages below 1%. Both A. millefolium essential oil and thujone exhibited a repellent activity against this insect at the higher doses tested (285.7 ng/cm(2)), demonstrating their potential as repellents for this species.

Morphology and Distribution of Sensilla on the Antennae of Hylamorpha elegans Burmeister (Coleoptera: Scarabaeidae).

Considering that sensilla constitute important functional elements of sensory systems in insects, the aim of this study was to determine the type and distribution of sensilla in the antennae of Hylamorpha elegans Burmeister examined by scanning electron microscopy. Hylamorpha elegans antennae are lamellate and consist of the scape, pedicel, and flagellum. The antennal club of this beetle consists of three terminal plates: proximal, middle, and distal lamellae. Four types of sensilla were observed in the lamellae from both sexes: sensilla trichoidea, chaetica, coeloconica, and placodea. Antennal length was larger in males than in females, and significant sexual variation in the number of sensilla placodea and sensilla coeloconica was observed.

Molecular Characterization and In Silico Analysis of the Pheromone-Binding Protein of the European Grapevine Moth Lobesia botrana (Denis & Schiffermüller) (Lepidoptera, Tortricidae).

The European grapevine moth Lobesia botrana (Denis & Schiffermüller) is an economically important insect in Europe. The species invaded vineyards in Chile, Argentina, and California during 2008-2010 causing severe problems. A major component of the sex pheromone, (E,Z)-7,9-dodecadienyl acetate (E7,Z9-12:Ac), is used in a mating disruption technique when grapevine moth populations are low or to monitor pest numbers. It is thought that these sexual pheromones are blends of volatiles that typically are specific to a species and are transported in the insect antenna by pheromone-binding proteins (PBPs) across the sensillar lymph to the olfactory receptors. Currently, an increasing number of Lepidopteran PBPs are being identified and cloned. However, there are no studies of the olfactory system and of proteins involved in the olfactory perception of L. botrana at the molecular level. In the present study, we report, for the first time, the sequence of a PBP from L. botrana (LbotPBP), which was determined using reverse transcription technology. Homology modeling was used to generate the three-dimensional protein structure. The model suggests that PBP consists of six α-helices as follows: Lys2-Met23 (α1), Thr28-Phe36 (α2), Arg46-Leu59 (α3), His70-Asn80 (α4), Glu84-Asn100 (α5), and Cys108-Lys125 (α6), held together by three disulfide bridges, Cys19-Cys54, Cys50-Cys108, and Cys97-Cys117. Docking simulations based on this model suggested that Trp114 is a key residue in the recognition of acetate pheromones, such as E7,Z9-12:Ac. In silico results in this study are consistent with previous findings in which E7,Z9-12:Ac acts as the most active compound in behavioral and electroantennographic assays.

Using volatile organic compounds to enhance atrazine biodegradation in a biobed system.

The effect of the terpenes α-pinene, eucalyptol, and limonene, individually and as mixtures, on atrazine (ATZ) biodegradation and on biological activity in a biobed biomixture was evaluated. Additionally, terpenes emitted from the biomixture were captured using solid-phase microextraction. Terpenes added individually at relatively low concentrations (50 µg kg(-1)) significantly enhanced ATZ degradation and biological activity during the first incubation days. No significant effect on ATZ degradation was found from adding the terpene mixture, and, interestingly, an inhibitory effect on phenoloxidase activity was found during the first 20 days of incubation when mixed terpenes were present at 100 µg kg(-1). Capturing terpenes demonstrated that during the first hour of incubation a significant fraction of the terpenes was volatilized. These results are the first to demonstrate the feasibility of using terpenes to enhance the degradation of a pesticide. However, successive applications of terpenes or the addition of materials that slowly release terpenes could sustain the ATZ degradation enhancement.

GA3 stimulates the formation and germination of somatic embryos and the expression of a KNOTTED-like homeobox gene of Cocos nucifera (L.).

The micropropagation of coconut palm has progressed rapidly; yet, there are constraints with regard to the number of somatic embryos formed and their germination. To overcome these, we tested the effect of gibberellic acid and characterized genes of the KNOX family. Gibberellic acid at 0.5 muM increased 1.5-fold the number of calli forming somatic embryos and twofold the number of somatic embryos per callus, calli with germinating embryos and the number of germinating somatic embryos per callus. With regard to the study of KNOX family genes, the complete sequences of two KNOX-like genes were obtained for CnKNOX1 and CnKNOX2. The deduced amino acid sequence of both showed highly conserved domains characteristic of KNOX genes. CnKNOX1 showed high homology with KNOX class I proteins. CnKNOX1 expression was detected throughout the embryogenesis process except in somatic embryos at the pro-globular stage, and was highest in somatic embryos at the coleoptilar stage. No detection of CnKNOX1 expression occurred in calli with aberrant embryos. The addition of gibberellic acid stimulated the expression of CnKNOX1 earlier and the relative expression at all stages was higher. CnKNOX2 expression occurred at all stages peaking at the globular stage, but gibberellic acid treatment decreased the expression. Gene expression was also analyzed in tissues of different organs of adult palms. With CnKNOX1, high level of expression was found in tissues of organs with, but not in those without, meristem, whereas CnKNOX2 expression was detected in tissues with and also in those without meristem.

Olfactory response of Haematobia irritans (Diptera: Muscidae) to cattle-derived volatile compounds.

In Chile, the horn fly, Hematobia irritans (L., 1758), is a major pest of grazing cattle and affects livestock production during the summer. Previous studies in Europe and the United States have shown that cattle flies, including H. irritans, are differentially attracted to individual cattle within herds and that volatile semiochemicals are responsible for this phenomenon. This study provides evidence that similar differential attractiveness occurs for the interaction between Chilean Holstein-Friesian cattle herds and local H. irritans populations. Thus, Holstein-Friesian dairy cattle, Bos taurus, which were of similar age and physiological condition, were shown to possess an uneven distribution of H. irritans. Heifers h6904 and h8104 were defined as low-carrier heifers and h5804, h2304 and h1404 as high-carrier heifers. Gas chromatography (GC) and coupled GC-mass spectrometric (GC-MS) analysis of samples collected from heifers revealed the presence of compounds previously reported as semiochemicals for cattle flies, including meta- and para-cresol, methylketones (C8-C11), and 6-methyl-5-hepten-2-one. Other compounds identified included carboxylic acids (butanoic, 3-methylbutanoic, pentanoic, and hexanoic acids), 1-hexanol, and 3-octanone. In Y-tube olfactometer studies, both m- and p-cresol attracted H. irritans at the highest doses tested (10(-6) g), as did the positive control 1-octen-3-ol. Of the other compounds tested, only 2-decanone and 2-undecanone produced a behavioral response, with significantly more flies being recorded in the control arm when the former compound was tested (at 10(-6) and 10(-8) g), and more flies being recorded in the treated arm for the latter compound (at 10(-7) g). This demonstration of behavioral activity with the identified compounds represents a first step for research into the application of semiochemicals in monitoring and control of cattle flies in Chile.

Insecticide resistance in the horn fly: alternative control strategies.

The horn fly, Haematobia irritans (Linnaeus 1758) (Diptera: Muscidae) is one of the most widespread and economically important pests of cattle. Although insecticides have been used for fly control, success has been limited because of the development of insecticide resistance in all countries where the horn fly is found. This problem, along with public pressure for insecticide-free food and the prohibitive cost of developing new classes of compounds, has driven the investigation of alternative control methods that minimize or avoid the use of insecticides. This review provides details of the economic impact of horn flies, existing insecticides used for horn fly control and resistance mechanisms. Current research on new methods of horn fly control based on resistant cattle selection, semiochemicals, biological control and vaccines is also discussed.

[Current immunologic and molecular data on the epidemiology of histoplasmosis in Morelos, Mexico]. / Actualidades inmunológicas y moleculares sobre la epidemiología de la histoplasmosis en Morelos, México.

The aim of the present paper was to analyze current data distribution of systemic mycosis, i.e. histoplasmosis, in the state of Morelos, Mexico. Data were collected based on immunoepidemiologic studies and preliminary molecular-epidemiologic results. The occupational activities of the rural population exposed to risk of infection, findings on genetic polymorphisms, and spread of the causative agent in nature were considered. These will be processed to propose criteria for establishing the prevalence of this mycosis in Morelos and to elaborate an epidemiologic map of the state.

[Efficacy of clopidogrel as ADP-dependent platelet aggregation inhibitor. Study on individuals with coronary artery disease]. / Eficacia del clopidogrel como inhibidor de la agregación plaquetaria dependiente de ADP. Un estudio en individuos con enfermedad arterial coronaria.

Acetyl-salicylic acid inhibits thromboxane A2 production and reduces the risk of vascular occlusive events by 20 to 25%. Ticlopidine inhibits ADP-dependent platelet aggregation and reduces the same risk by 30 to 35%, but produces some adverse effects. Clopidogrel is a ticlopidin-derived antiplatelet-drug, with the same mechanism of action; reduces the expression of the glycoprotein IIb/IIIa, the fibrinogen receptor on the platelet surface. Clopidogrel has the same clinical efficacy of ticlopidin and lowers the incidence of adverse effects. In this study, we evaluated the effects of one daily dosis of 75 mg of clopidogrel on platelet function in 33 subjects with coronary artery disease. Before treatment and after the 6th and 12th week, the following parameters were evaluated: 5 microM-ADP and 20 micrograms/mL collagen-induced platelet aggregation, bleeding time and fibrinogen concentration. In basal and in the 6th and 12th week samples, ADP-induced platelet aggregation was 90.7% +/- 13.2, 54.6% +/- 23.2 and 49.2% +/- 23.7 respectively, that represents a significant reduction of 38.6% and 44.4%. Reduction of collagen-induced platelet aggregation was not significative. Plasmatic fibrinogen did not suffer variation during treatment. Bleeding time was significant prolonged from 4.1 minutes to 15.4 and 14.6 minutes (3.7-3.5 times compared with the test before treatment). There were no haemorrhagic complications, only digestive discomfort in fewer than 3% of patients. We concluded that clopidogrel is a safe and efficacious drug for patients, it efficiently reduces ADP-induced platelet aggregation and prolongs bleeding time.

[Investigation of HBV-DNA using the polymerase chain reaction (PCR) in HBsAg-negative, anti-HBc-positive Venezuelan donors]. / Investigación del VHB-ADN por reacción en cadena de la polimerasa (RCP) en donantes venezolanos AgHBs negativos/Anti HBc positivos.

OBJECTIVE: In our Center the disposal of blood with HBV positive markers is approximately 6%, being 90% AgsHB negative and anti HBc positive. With the purpose of knowing the infected capacities of these donations and to consider the possibility of using them for transfusion, the presence of the viral genoma was investigated by PCR, in a group of samples with these characteristics that were also anti sHB positive. They were correlated with the readings of the anti cHB total, with the anti cHB-IgM and with the titration of the anti sHB. MATERIALS AND METHODS: 87/100 random samples, from February to June 1996, were frozen at -30 degrees C for their later evaluation. In the serological screening were used Auszyme Monoclonal of Abbott and the Heprofile anti cHB, ADI-Diagnostic. The readings of the anti cHB was considered strong, moderate or weak according to its distance to the cut off. For the determination of the anti sHB, Hepanostika anti sHB, Organon Technika was used. They were considered with low titers (< 10 UI/L), high (> or = 10 UI/L) and very high (> or = 100 UI/L). In the determination of the anti cHB-IgM, Heprofile ADI-Diagnostic was used. For the investigation of the viral genoma, it was carried out with double PCR-ADN, using two internal and two external primers for the core-precore region. RESULTS: 70/87 (80.45%) of the samples presented high readings of anti cHB with high titers of anti sHB, being positive for anti cHB-IgM seven of them and one was HBV-ADN positive (1.42%) suggesting the possibility to be cronic carrier. In the remaining 19.55% of the samples we didn't detect positive results in the amplification assays. CONCLUSIONS: 1. The donors showed high levels of immunocompetence. 2. High titer of anti sHB doesn't guarantee the absence of viral genoma and therefore the absence of infectivity can not be sure. This doesn't allow us to come in like donors neither to use their blood in the transfusional therapy.

[Serum hepatitis B markers in blood donors in Venezuela. What do they mean?]. / Marcadores serológicos para hepatitis B (HB) en donantes venezolanos qué significan?

PURPOSE: The prevalence of serum BH markers is high in our country. In order to assess the factors of impact in this fact a group of blood donors with positive BH markers attending our Service between 1-1-95 and 12-31-96 was evaluated, although the endemic character of the disease may be an obstacle for its recognition. The residual risk of HBV transmission by seronegative transfusions was also evaluated. MATERIAL AND METHODS: The evaluation of donors was performed by surveying identification data, type and frequency of donations and risk factors. The following reagents were used to carry out the bests: Biotest Elisa HbsAgand Heprofile anti HBc, ADI Diagnostics (1995), and Abbott Auzyme Monoclonal and Abbott Corzyme Diagnostic kit. Positive donors were re-evaluated to determine any progression of the disease and they were classified into false positive (FP), chronic carriers (CC), acute hepatitis (AH), naturally immunized (NI), or without evidence of immunity (WI). The residual transfusional risk was calculated by multiplying the adjusted prevalence of HbsAg positive donors by the window period expressed in fractions of a year. RESULTS: Out of 53,338 donations, 5.91% were discarded due to the presence of HBV, 7.5% of them showing significant association with other markers: HCV, Chagas' disease and syphilis. Of all donors 223 (7.54%) returned for re-evaluation, and only 139 (62.33% of them) could be fully evaluated as follows: FP 10.09%, AH 5.76%, CC 7.19%, NI 68.35%, WI 7.91%. Their mean age was 35.73 +/- 9.95 years, and there were 112 males and 12 females. The donations were patient-related in 93.5% of cases, voluntary in 3.22% and coming from mobile units in 3.22% of instances (p = 0.067). Fifth per cent donated for the first time, and the other 50% had donated previously although not in a consecutive way (p = 0.96): 46% were ignorant of their post-donation findings, 37.09% were seemingly negative and 16.12% had positive tests. Of these last, the reasons for donation included defective questioning and information (50%), results not verified (20%) and family/hospital pressure (30%). Homo/bisexuality and drug abuase (not intravenous) were found significant when associated to each other or to promiscuity. Promiscuity attained significant risk value, especially when previous venereal disease or close contact with BH patients were presented (p = 0.0015, p = 0.155, and p = 0.0036, respectively). The residual risk of transmitting HBV by seronegative transfusions was found to be: 1:437. CONCLUSIONS: 1. No risk factors were detected in a half of the cases. 2. The other half, comprising the known seropositive subjects and the promiscuous, homo/bisexual donors and those having close contact with HBV patients, can be discarded predonation. 3. Drug abuse (not intravenous), frequently associated with other risk factors, should discard a donor. 4. History of veneral disease should be considered a risk factor due to its common association to HBV. 5. The residual risk of acquiring HBV post-transfusion is still high. RECOMMENDATIONS: 1. To improve the information provided to donors regarding the risk factors. 2. The interrogation should be simple, direct and deep. 3. To insist in the importance of the donor returning for his/her credentials in the blood bank and to know about any altered results, and to seek medical advice when appropriate. 4. To change related donation into a voluntary and consecutive pattern.

[Tetralogy of Fallot in adults]. / Tetralogía de Fallot en adultos.

We present the clinical and hemodynamic profile of 33 patients, older than the 18 year with tetralogy of Fallot. Cardiac catheterization and selective angiocardiography were performed in all cases. We excluded patients with valvular pulmonary atresia associated with tetralogy of Fallot. Most of the patients were in functional class II or IV of the New York Heart Association and all presented with cyanosis. In the electrocardiogram of 23 patients we found right bundle branch block. None had significant cardiomegaly, 19 of 20 cases had reduced pulmonary blood flow and reticular pattern in the lung fields as observed in the chest X-ray. We did not find correlation between the age and the degree of cyanosis, but the level of arterial desaturation correlated with the functional class. The right ventricular systolic pressure was elevated in all patients. In all cases but one the pulmonary arterial systolic pressure were normal of slightly increased. One case with mild pulmonary infundibular stenosis had pulmonary systolic pressure similar to the systemic pressure. Multivariate analysis for functional class showed significant value for cyanosis. Systemic-pulmonary shunt was performed in 10 patients with hypoplastic pulmonary arteries plus reduced pulmonary blood flow, with one postoperative death. We did not find postoperative mortality in patients who underwent corrective surgery. The anatomic and functional behavior of patients who underwent surgery, allowed better tolerance to their heart defects.