Changes in the Neurochemical Coding of the Anterior Pelvic Ganglion Neurons Supplying the Male Pig Urinary Bladder Trigone after One-Sided Axotomy of Their Nerve Fibers

The present study investigated the effect of unilateral axotomy of urinary bladder trigone (UBT)-projecting nerve fibers from the right anterior pelvic ganglion (APG) on changes in the chemical coding of their neuronal bodies. The study was performed using male pigs with immunohistochemistry and quantitative real-time PCR (qPCR). The animals were divided into a control (C), a morphological (MG) or a molecular biology group (MBG). APG neurons supplying UBT were revealed using the retrograde tracing technique with Fast Blue (FB). Unilateral axotomy resulted in an over 50% decrease in the number of FB+ neurons in both APG ganglia. Immunohistochemistry revealed significant changes in the chemical coding of FB+ cells only in the right ganglion: decreased expression of dopamine-B-hydroxylase (DBH)/tyrosine hydroxylase (TH) and up-regulation of the vesicular acetylcholine transporter (VAChT)/choline acetyltransferase (ChAT), galanin (GAL), vasoactive intestinal polypeptide (VIP) and brain nitric oxide synthase (bNOS). The qPCR results partly corresponded with immunofluorescence findings. In the APGs, genes for VAChT and ChAT, TH and DBH, VIP, and NOS were distinctly down-regulated, while the expression of GAL was up-regulated. Such data may be the basis for further studies concerning the plasticity of these ganglia under experimental or pathological conditions.

Age Dependency of the Prognostic Impact of Tumor Genomics in Localized Resectable MYCN-Nonamplified Neuroblastomas. Report From the SIOPEN Biology Group on the LNESG Trials and a COG Validation Group

PURPOSE For localized, resectable neuroblastoma without MYCN amplification, surgery only is recommended even if incomplete. However, it is not known whether the genomic background of these tumors may influence outcome. PATIENTS AND METHODS Diagnostic samples were obtained from 317 tumors, International Neuroblastoma Staging System stages 1/2A/2B, from 3 cohorts: Localized Neuroblastoma European Study Group I/II and Children’s Oncology Group. Genomic data were analyzed using multi- and pangenomic techniques and fluorescence in-situ hybridization in 2 age groups (cutoff age, 18 months) and were quality controlled by the International Society of Pediatric Oncology European Neuroblastoma (SIOPEN) Biology Group. RESULTS Patients with stage 1 tumors had an excellent outcome (5-year event-free survival [EFS] ± standard deviation [SD], 95% ± 2%; 5-year overall survival [OS], 99% ± 1%). In contrast, patients with stage 2 tumors had a reduced EFS in both age groups (5-year EFS ± SD, 84% ± 3% in patients < 18 months of age and 75% ± 7% in patients ≥ 18 months of age). However, OS was significantly decreased only in the latter group (5-year OS ± SD in < 18months and ≥ 18months, 96% ± 2% and 81% ± 7%, respectively; P = .001). In < 18months, relapses occurred independent of segmental chromosome aberrations (SCAs); only 1p loss decreased EFS (5-year EFS ± SD in patients 1p loss and no 1p loss, 62% ± 13% and 87% ± 3%, respectively; P = .019) but not OS (5-year OS ± SD, 92% ± 8% and 97% ± 2%, respectively). In patients ≥ 18 months, only SCAs led to relapse and death, with 11q loss as the strongest marker (11q loss and no 11q loss: 5-year EFS ± SD, 48% ± 16% and 85% ± 7%, P = .033; 5-year OS ± SD, 46% ± 22% and 92% ± 6%, P = .038). CONCLUSION Genomic aberrations of resectable non– MYCN-amplified stage 2 neuroblastomas have a distinct age-dependent prognostic impact. Chromosome 1p loss is a risk factor for relapse but not for diminished OS in patients < 18 months, SCAs (especially 11q loss) are risk factors for reduced EFS and OS in those > 18months. In older patients with SCA, a randomized trial of postoperative chemotherapy compared with observation alone may be indicated.

Structural investigation of cyclic nucleotide binding proteins from Trypanosoma cruzi

Fora targeted therapy of Trypanosomiasis, new antiparasitic drugs should be specifically directed against essential pathways in the parasite life cycle. Among these potential targets are signal transduction pathways, which have remained largely unexplored in Trypanosoma species. Of special interest is cAMP-mediated signaling, since cAMP has been shown to play critical roles in the life cycle of T. cruzi and in host cell during invasion. The presented research focuses on the identification and characterisation of novel cAMP response proteins (CARPs) in T. cruzi by using a multi disciplinary approach involving the parasitology group of Dr Martin Edreira (University of Buenos Aires, Argentina) and the structural biology group of Dr Ivan Campeotto (University of Leicester, UK). The aim of the project is not only to increase our knowledge about T. cruzi biology but also to target CARPs for the design and development of novel therapeutic agents against Chagas disease. To date, protein crystals of one of the members of the CARP family have been obtained, paving the way for structure determination and for a structure-based drug design approach.

Effect of the Technology-Supported Learning on the Academic Performance of Secondary School Students

A Posttest only Controlled Group Experiment research was used to explore the effect of teaching through the technology- supported environment on learners� performance. Students of Biology group Grade IX, (n = 177), from public schools of Karachi, participated in the experimental research for 20 weeks in the year 2016-17. The technologysupported learning was intervened in the experiment group whereas, a conventional teaching was intervened in the controlled group. The annual examination�s 2017 result was examined and scores of both experimental and controlled participants were compared by the independent sample t-test. The data analysis showed a noteworthy variance in the scores of controlled and experiment batch. The students studying in a technology-supported

DEHIDRASI DAN PEMBEKUAN JARINGAN APEKS TEBU UNTUK PENYIMPANAN JANGKA PANJANG

<p>ABSTRAK<br />Tebu (Saccharum officinarum L.) merupakan tanaman yang<br />diperbanyak secara vegetatif. Kriopreservasi merupakan metode yang<br />paling sesuai untuk penyimpanan jangka panjang bagi tanaman yang<br />diperbanyak secara vegetatif. Dehidrasi dan pembekuan jaringan merupa-<br />kan tahapan paling kritis yang menentukan keberhasilan kriopreservasi.<br />Tujuan penelitian adalah untuk memperoleh durasi dehidrasi yang optimal<br />dan metode pembekuan jaringan apeks tebu. Penelitian dilakukan di<br />Laboratorium Kultur Jaringan, Kelompok Peneliti Biologi Sel dan<br />Jaringan, Balai Penelitian dan Pengembangan Bioteknologi dan Sumber-<br />daya Genetik Pertanian, Bogor pada Mei 2013 sampai Februari 2014.<br />Untuk optimasi metode dehidrasi, apeks direndam dalam larutan PVS2<br />(MS + gliserol 30% + etilen glikol 15% + dimetil sulfoksida 15% +<br />sukrosa 0,4 M) selama 10, 20, 30, dan 40 menit. Untuk optimasi metode<br />pembekuan, diujikan kombinasi perlakuan prakultur (dengan sukrosa 0;<br />0,1; dan 0,3 M selama 5 hari) dan pemuatan dalam larutan LS (MS +<br />gliserol 2 M + sukrosa 0,4 M) selama 0, 10, 20, dan 30 menit sebelum<br />tahapan dehidrasi dan pembekuan jaringan di dalam nitrogen cair (-<br />196 o C). Hasil penelitian menunjukkan durasi dehidrasi jaringan yang<br />terbaik adalah 30 menit dalam larutan PVS2. Kombinasi perlakuan<br />prakultur dengan sukrosa 0,3 M dan pemuatan dengan larutan LS selama<br />10 menit merupakan metode terbaik untuk pembekuan jaringan. Persentase<br />tumbuh sebelum dan setelah pembekuan dalam nitrogen cair berturut-turut<br />adalah 100 dan 40%. Setelah kriopreservasi, biakan mampu tumbuh<br />dengan tingkat multiplikasi tunas sekitar 10 tunas/eksplan. Metode yang<br />diperoleh pada penelitian ini berpeluang diterapkan untuk penyimpanan<br />plasma nutfah tebu dalam jangka panjang secara kriopreservasi dan<br />eliminasi patogen obligat secara krioterapi.<br />Kata kunci: Saccharum officinarum L., apeks, dehidrasi, pembekuan,<br />nitrogen cair</p><p>ABSTRACT<br />Sugarcane (Saccharum officinarum L.) is vegetatively propagated<br />plant. Cryopreservation is the most suitable method for long-term<br />preservation of vegetatively propagated plant. Dehydration and freezing<br />are critical steps of successful cryopreservation so that it should be<br />optimized. The research aimed to obtain the optimal duration of<br />dehydration and freezing method of sugarcane apex tissues. The<br />experiments were conducted at Tissue Culture Laboratory, Plant Cell<br />Tissue Biology Group, Indonesian Center for Agricultural Biotechnology<br />and Genetic Resources Research and Development on May<br />2013−February 2014. To optimize dehydration method, the tissues were<br />exposured in PVS2 solution (MS + 30% glycerol + 15% ethylene glycol +<br />15% dimethyl sulphoxide + 0.4 M sucrose) for 10, 20, 30, and 40 minutes.<br />To optimize freezing method, the combined treatment of preculture with<br />sucrose (0, 0.1, dan 0.3 M) for 5 days and loading in LS solution (MS + 2<br />M glycerol + 0.4 M sucrose) for 0, 10, 20, dan 30 minutes) were tested<br />before dehydration for 30 minutes and freezing in liquid nitrogen (-196 o C).<br />The best duration of dehydration was 30 minutes. The combined treatment<br />of preculture on 0.3 M sucrose and loading for 10 minutes was the best<br />method for tissues freezing. Percentage of regrowth before and after<br />freezing in liquid nitrogen was 100 and 40% respectively. After<br />cryopreservation, the cultures could grow with high shoot multiplication<br />rate about 10 shoots/explant. The method resulted in this study can be<br />applied for long-term storage of sugarcane germplasms by cryopreser-<br />vation and (elimination of obligate pathogens by cryotherapy.<br />Keywords: Saccharum officinarum L., apex, dehydration, freezing, liquid<br />nitrogen.</p>

In crystallooptical spectroscopy (icOS) as a complementary tool on the macromolecular crystallography beamlines of the ESRF

The analysis of structural data obtained by X-ray crystallography benefits from information obtained from complementary techniques, especially as applied to the crystals themselves. As a consequence, optical spectroscopies in structural biology have become instrumental in assessing the relevance and context of many crystallographic results. Since the year 2000, it has been possible to record such data adjacent to, or directly on, the Structural Biology Group beamlines of the ESRF. A core laboratory featuring various spectrometers, named the Cryobench, is now in its third version and houses portable devices that can be directly mounted on beamlines. This paper reports the current status of the Cryobench, which is now located on the MAD beamline ID29 and is thus called the ID29S-Cryobench (where S stands for `spectroscopy'). It also reviews the diverse experiments that can be performed at the Cryobench, highlighting the various scientific questions that can be addressed.

In crystallo optical spectroscopy as a complementary tool to crystallography

The analysis of structural data obtained by X-ray crystallography benefits from information obtained from complementary techniques, especially if these are applied to the crystals themselves. As a consequence, optical spectroscopies as applied in Structural Biology have become instrumental in assessing the relevance of many crystallographic results. Since the year 2000, such data can be recorded close to, or directly on, the Structural Biology Group beamlines of the ESRF. A core laboratory featuring various spectrometers, named the Cryobench, is now in its third version and houses portable devices that can be directly mounted on beamlines. This presentation will report the status of the current version of the Cryobench, now located on the MAD beamline ID29 and thus called ID29S-Cryobench, S standing for 'Spectroscopy'. In particular, the new on-line Raman data collection mode of ID29 will be described. Finally, it will review the diverse experiments that can be performed at the Cryobench, highlighting various scientific questions that can be addressed.

Chronic Myeloid Leukemia (CML) Patients With Atypical e1a2 P190 BCR-ABL Translocation Show a Poor Response To Therapy With Tyrosine Kinase Inhibitors (TKI)

Introduction P210 BCR-ABL translocation resulting from rearrangements within the major breakpoint cluster region (M-BCR), either e13a2 or e14a2, is the molecular hallmark of chronic myeloid leukemia (CML). However, some CML patients may harbor atypical BCR-ABL rearrangements such e1a2 P190 BCR-ABL which involves the minor breakpoint cluster region (m-BCR). Response to therapy with tyrosine kinase inhibitors (TKI) and outcome of such atypical patients is not well defined. Objective To evaluate response to TKI therapy of CML patients with the atypical e1a2 P190 BCR-ABL translocation. Patients and Methods Since 2009, 4 patients with CML in chronic phase and with atypical e1a2 P190 BCR-ABL rearrangement have been recruited in various institutions belonging to the Hematological Molecular Biology Group (GBMH) of the Spanish Society of Hematology (SEHH). Patient characteristics, treatments administered and response to therapy for the 4 patients is shown in Table 1. BCR-ABL transcripts were revealed at diagnosis by quantitative PCR followed by conventional agarose electrophoresis of PCR products. Molecular follow-up of BCR-ABL transcripts throughout treatment was performed by quantitative PCR following the guidelines of the European Leukemia Net. Results One patient received treatment (HU and INF+araC) prior to TKI (Pat. 1; Table 1). All 4 patients received Imatinib as initial TKI treatment. Two of the patients treated with Imatinib (Pat. 1,2) obtained a complete molecular response (CMR) and the other 2 (Pat. 3,4) only achieved a complete hematological response (CHR) as best response (Table 1). All patients had to switch to a second generation TKI (3 Nilotinib and 1 Dasatinib) due to intolerance to Imatinib (n=1; Pat. 1) or resistance (n=3; Pat. 2-4). The patient who received Dasatinib as second line TKI (Pat. 3) only achieved a partial hematologic response (PHR) and was changed to Nilotinib as third line TKI, achieving CHR after which the patient entered in blast crisis and died 36 months after diagnosis (Table 1). Overall, only 1 (Pat. 1) out of the 4 patients included in the present study achieved a sustained molecular response with Imatinib. At last follow-up, among the 4 patients included in the study, all 4 had needed a change of TKI, 1 had died due to disease progression (Pat. 3) and only 2 of them retained a molecular response (Pat. 1,2). Conclusion CML patients harboring atypical e1a2 P190 BCR-ABL transcripts show a poor response and short-lived responses to TKI therapy and therefore should be identified as high-risk patients at diagnosis. These patients must be closely monitored during therapy with TKI and should be treated upfront with a second generation TKI or even be considered for allogeneic SCT in the early phase of the disease. Paper presented on behalf of the Hematological Molecular Biology Group (GBMH) of the Spanish Society of Hematology (SEHH). AJ-V and IB contributed equally to this work. Disclosures: No relevant conflicts of interest to declare.