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      Volume 30, Issue 5

      December 2005,   pages  551-795

    • Clipboard: Tumour development due to stroma permissiveness

      Edna Cukierman

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    • Clipboard: Cellular clocks: circadian rhythms in primary human fibroblasts

      Martha Merrow Cornelia Boesl Till Roenneberg

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    • Commentary: How relevant are the concepts of species diversity and species richness?

      Laishangbam Sanjit Dinesh Bhatt

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    • Commentary: Neuronal survival in epilepsy: to die or not to die?

      Subramaniam Ganesh Shweta Singh

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    • Commentary: Whither genome research: Of man, mosquito and malaria

      Utpal Tatu Samta Jain P Padma Priya

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    • N V Timofeeff-Ressovsky in Germany (July, 1925–September, 1945)

      Yakov G Rokityanskij

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    • Living in a physical world V. Maintaining temperature

      Steven Vogel

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    • What history tells us III. André Lwoff: From protozoology to molecular definition of viruses

      Michel Morange

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    • Association of genetic polymorphism in GH gene with milk production traits in Beijing Holstein cows

      Guo Li Zhou Hai Guo Jin Chen Liu Shan Li Qi Zhu Yu Hou Wu

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      Associations were analysed between polymorphisms of the growth hormone gene (GH-MspI) (localized in intron 3) and milk production traits of Beijing Holstein cows (a total of 543 cows). Polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method was used for identification of various genotypes. Frequencies of genotypes were 0.77, 0.21 and 0.02 for A/A, A/B and B/B, respectively. The frequency of the 𝐺𝐻𝐴 allele is 0.875.

      The results of the least squares means show that in all three lactations, the GH A/A cows yielded more milk (𝑃 < 0.01 for lactation I and 𝑃 < 0.05 for lactations II and III), whereas A/B cows showed higher milk fat content than A/A individuals (𝑃 < 0.05 for lactations I and II, and 𝑃 < 0.01 for lactation III). The A/A cows yielded more fat than A/B individuals (𝑃 < 0.01 only in lactation I). The A/A cows yielded more milk protein than A/B individuals (𝑃 < 0.01 for lactations I, II, and III). The A/A cows produced milk of higher protein content than of A/B individuals (𝑃 < 0.05 only in lactation II).

    • Single gene retrieval from thermally degraded DNA

      Lianwen Zhang Lianwen Zhang

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      To simulate single gene retrieval from ancient DNA, several related factors have been investigated. By monitoring a 889 bp polymerase chain reaction (PCR) product and genomic DNA degradation, we find that heat and oxygen (especially heat) are both crucial factors influencing DNA degradation. The heat influence, mainly represented by temperature and heating time, affects the DNA degradation via DNA depurination followed by cleavage of nearby phosphodiesters. The heating time influence is temperature-dependent. By reactive oxygen species (ROS) scavenging and 1,3-diphenyl-isobenzofuran (DPBF) bleaching experiments the influence of oxygen on DNA thermal degradation was shown to occur via a singlet oxygen pathway. A comparative study of the thermal degradation of cellular DNA and isolated DNA showed that cellular lipids can aggravate DNA thermal degradation. These results confirm the possibility of gene amplification from thermally degraded DNA. They can be used to evaluate the feasibility of the retrieval of single gene from ancient remains.

    • Cloning and sequence analysis of the Antheraea pernyi nucleopolyhedrovirus gp64 gene

      Wenbing Wang Shanying Zhu Liqun Wang Feng Yu Weide Shen

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      Frequent outbreaks of the purulence disease of Chinese oak silkworm are reported in Middle and Northeast China. The disease is produced by the pathogen Antheraea pernyi nucleopolyhedrovirus (AnpeNPV). To obtain molecular information of the virus, the polyhedra of AnpeNPV were purified and characterized. The genomic DNA of AnpeNPV was extracted and digested with HindIII. The genome size of AnpeNPV is estimated at 128 kb. Based on the analysis of DNA fragments digested with HindIII, 23 fragments were bigger than 564 bp. A genomic library was generated using HindIII and the positive clones were sequenced and analysed. The gp64 gene, encoding the baculovirus envelope protein GP64, was found in an insert. The nucleotide sequence analysis indicated that the AnpeNPV gp64 gene consists of a 1530 nucleotide open reading frame (ORF), encoding a protein of 509 amino acids. Of the eight gp64 homologues, the AnpeNPV gp64 ORF shared the most sequence similarity with the gp64 gene of Anticarsia gemmatalis NPV, but not Bombyx mori NPV. The upstream region of the AnpeNPV gp64 ORF encoded the conserved transcriptional elements for early and late stage of the viral infection cycle. These results indicated that AnpeNPV belongs to group I NPV and was far removed in molecular phylogeny from the BmNPV.

    • The amino-terminal domain of human signal transducers and activators of transcription 1: Overexpression, purification and characterization

      Arati Prabhu Evans Coutinho Sudha Srivastava

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      The dual functional signal transducers and activators of transcription (STAT) proteins are latent cytoplasmic transcription factors that play crucial roles in host defense. Animals that lack these proteins are highly susceptible to microbial and viral infections and chemically induced primary tumours. We have over expressed the aminoterminal domain of human STAT1 (hSTAT1) in Escherichia coli and purified it by affinity chromatography and gel filtration chromatography. The entire process has been monitored by gel electrophoresis. The pure protein has been characterized by mass spectrometry and 2-dimensional nuclear magnetic resonance (2D-NMR) spectroscopy. Our results indicate that the N-terminus of hSTAT1 exists as a dimer in solution.

    • Molecular phylogenetic analysis of Vibrio cholerae O1 El Tor strains isolated before, during and after the O139 outbreak based on the intergenomic heterogeneity of the 16S-23S rRNA intergenic spacer regions

      Atreyi Ghatak Anasuya Majumdar Ranajit K Ghosh

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      We have cloned, sequenced and analysed all the five classes of the intergenic (16S-23S rRNA) spacer region (ISR) associated with the eight rrn operons (rrna-rrnh) of Vibrio cholerae serogroup O1 El Tor strains isolated before, during and after the O139 outbreak. ISR classes ‘a’ and ‘g’ were found to be invariant, ISR-B (ISRb and ISRe) exhibited very little variation, whereas ISR-C (ISRc, ISRd, and ISRf) and ISRh showed the maximum variation. Phylogenetic analysis conducted with all three ISR classes (ISR-B, ISR-C and ISRh) showed that the pre-O139 serogroup and post-O139 serogroup O1 El Tor strains arose out of two independent clones, which was congruent with the observation made by earlier workers suggesting that analyses of ISR-C and ISR-h, instead of all five ISR classes, could be successfully used to study phylogeny in this organism.

    • Enhancement of resistance to aphids by introducing the snowdrop lectin gene gna into maize plants

      Zhaoyu Wang Kewei Zhang Xiaofen Sun Kexuan Tang Juren Zhang

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      In order to enhance the resistance to pests, transgenic maize (Zea mays L.) plants from elite inbred lines containing the gene encoding snowdrop lectin (Galanthus nivalis L. agglutinin; GNA) under control of a phloemspecific promoter were generated through the Agrobacterium tumefaciens-mediated method. The toxicity of GNA-expressing plants to aphids has also been studied. The independently derived plants were subjected to molecular analyses. Polymerase chain reaction (PCR) and Southern blot analyses confirmed that the gna gene was integrated into maize genome and inherited to the following generations. The typical Mendelian patterns of inheritance occurred in most cases. The level of GNA expression at 0.13%–0.28% of total soluble protein was observed in different transgenic plants. The progeny of nine GNA-expressing independent transformants that were derived separately from the elite inbred lines DH4866, DH9942, and 8902, were selected for examination of resistance to aphids. These plants synthesized GNA at levels above 0.22% total soluble protein, and enhanced resistance to aphids was demonstrated by exposing the plants to corn leaf aphid (Rhopalosiphum maidis Fitch) under greenhouse conditions. The nymph production was significantly reduced by 46.9% on GNA-expressing plants. Field evaluation of the transgenic plants supported the results from the inoculation trial. After a series of artificial self-crosses, some homozygous transgenic maize lines expressing GNA were obtained. In the present study, we have obtained new insect-resistant maize material for further breeding work.

    • Biochemical characterization of N-methyl N' -nitro-N-nitrosoguanidine-induced cadmium resistant mutants of Aspergillus niger

      Samar Kumar Pal Tapan Kumar Das

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      Two cadmium resistant mutants (Cd1 and Cd2) of Aspergillus niger, among the six isolated by mutagenization with N-methyl N′-nitro-N-nitrosoguanidine (MNNG) at pH 6.4 were selected for the study. Analysis of lipid composition of the mutants and the wildtype indicated that total lipid as well as individual lipids of the cadmium resistant mutants were changed as compared with that of the wildtype. The increased activities of metal-lothionein and reduced activities of D-xylose isomerase and L-phenylalanine ammonia lyase in cell free extract of the cadmium resistant mutants suggested that mutants could allow high concentration of cadmium salt as compared with that of the wildtype. The respiratory activity and intracellular as well as extracellular Cd2+ concentration of the mutants reflected the high tolerance of the Cd mutants to cadmium ion.

    • Transgenic tobacco plants expressing BoRS1 gene from Brassica oleracea var. acephala show enhanced tolerance to water stress

      Dongqin Tang Hongmei Qian Lingxia Zhao Danfeng Huang Kexuan Tang

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      Water stress is by far the leading environmental stress limiting crop yields worldwide. Genetic engineering techniques hold great promise for developing crop cultivars with high tolerance to water stress. In this study, the Brassica oleracea var. acephala BoRS1 gene was transferred into tobacco through Agrobacterium-mediated leaf disc transformation. The transgenic status and transgene expression of the transgenic plants was confirmed by polymerase chain reaction (PCR) analysis, Southern hybridization and semi-quantitative one step RT-PCR analysis respectively. Subsequently, the growth status under water stress, and physiological responses to water stress of transgenic tobacco were studied. The results showed that the transgenic plants exhibited better growth status under water stress condition compared to the untransformed control plants. In physiological assessment of water tolerance, transgenic plants showed more dry matter accumulation and maintained significantly higher levels of leaf chlorophyll content along with increasing levels of water stress than the untransformed control plants. This study shows that BoRS1 is a candidate gene in the engineering of crops for enhanced water stress tolerance.

    • Expression profiles of hot pepper (Capsicum annuum) genes under cold stress conditions

      Eul-Won Hwang Kyung-A Kim Soo-Chul Park Mi-Jeong Jeong Myung-Ok Byun Hawk-Bin Kwon

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      In an attempt to determine a cold defense mechanism in plants, we have attempted to characterize changes occurring in the expression of cold-regulated transcript levels in the hot pepper (Capsicum annuum), using cDNA microarray analysis, combined with Northern blot analysis. After analysing a 3.1 K hot pepper cDNA microarray, we isolated a total of 317 cold inducible genes. We selected 42 genes which were up-regulated and three genes which were down-regulated due to cold treatment, for further analysis. Among the 45 genes which appeared to be up-regulated by cold, 19 genes appeared to be simultaneously regulated by salt stress. Among the up-regulated cold-stress genes, we identified a variety of transcription factors, including: a family of 4 ethylene-responsive element binding protein (EREBP, designated CaEREBP-C1 to C4) genes, a bZIP protein (CaBZ1), RVA1, Ring domain protein, HSF1, and the WRKY (CaWRKY1) protein. As mentioned earlier, several genes appeared to be induced not only by cold stress, but also simultaneously by salt stress. These genes included: CaEREBP-C3, CaBZ1, putative trans-activator factor, NtPRp27, malate dehydrogenase, putative auxin-repressed protein, protein phosphatase (CaTPP1), SAR8.2 protein precursor, late-embryogenesis abundant protein 5 (LEA5), DNAJ protein homologue, xyloglucanendo-1,4-𝛽-D-gucanase precursor, PR10, and the putative non-specific lipid transfer protein StnsLTP.

    • Single primer amplification reaction methods reveal exotic and indigenous mulberry varieties are similarly diverse

      Esha Bhattacharya S B Dandin Shirish Anand Ranade

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      Mulberry is the sole food source for mulberry silkworm and a number of indigenous and exotic varieties are used in sericulture. Studies on assessment of genetic diversity have been done amongst a few mulberry varieties using one or at the most two methods. However, no comprehensive study on a large number of varieties has been carried out. In present study, single primer amplification reaction (SPAR) methods have been used for determination of diversity in 27 mulberry varieties (exotic as well as indigenous), using four minisatellite core sequence primers for directed amplification of minisatellite DNA (DAMD), three simple sequence repeat (SSR) motifs as primers for inter simple sequence repeat (ISSR) and 20 arbitrary sequence decamer primers for random amplified polymorphic DNA (RAPD) reactions. The Jaccard coefficients were determined for the DAMD, ISSR and RAPD band data (total of 58, 39 and 235 bands respectively). All three methods revealed wide range of distances supporting a wide range of mulberry genetic diversity. A cumulative analysis of the data generated by three methods resulted in a neighbour-joining (NJ) tree that gave a better reflection of the relatedness and affinities of the varieties to each other. Comparison of the three methods by marker indices and the Mantel test of correlation indicated that though all methods were useful for the assessment of diversity in mulberry, the DAMD method was better. When considered as two groups (10 exotic and 17 indigenous varieties), the mulberry varieties in the exotic group were found to have slightly greater diversity than the indigenous ones. These results support the concept of naturalization of mulberry varieties at locales distant from their origins.

    • Chemosensory processing in the fruit fly, Drosophila melanogaster: Generalization of a feeding response reveals overlapping odour representations

      Sathees B C Chandra Sandeep Singh

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      Insects are capable of detecting, and discriminating between, a very large number of odours. The biological relevance of many of those odours, particularly those related to food, must first be learned. Given that the number of sensory receptors and antennal lobe (AL) glomeruli is limited relative to the number of odours that must be detectable, this ability implies that the olfactory system makes use of a combinatorial coding scheme whereby each sensory cell or AL projection neuron can participate in coding for several different odours. An important step in understanding this coding scheme is to behaviourally quantify the degree to which sets of odours are discriminable. Here we evaluate odour discriminability in the fruit fly, Drosophila melanogaster, by first conditioning individual flies to not respond to any of several odourants using a nonassociative conditioning protocol (habituation). We show that flies habituate unconditioned leg movement responses to both mechanosensory and olfactory stimulation over 25 unreinforceed trials. Habituation is retained for at least 2 h and is subject to dishabituation. Finally, we test the degree to which the conditioned response generalizes to other odourants based on molecular features of the odourants (e.g. carbon chain length and the presence of a target functional group). These tests reveal predictable generalization gradients across these molecular features. These data substantiate the claim that these features are relevant coding dimensions in the fruit fly olfactory system, as has been shown for other insect and vertebrate species.

    • Phenotypic plasticity and reaction norms of abdominal bristle number in Drosophila melanogaster

      Brigitte Moreteau Jean R David

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      The phenotypic plasticity of abdominal bristle number (segments 3 and 4 in females) was investigated in 10 isofemale lines from a French population, grown at 7 constant temperatures, ranging from 12° to 31°C. Overall concave reaction norms were obtained with a maximum around 20°–21°C. Intraclass correlation (isofemale line heritability) was not affected by temperature. Correlations between segments 3 and 4 strongly contrasted a low within-line phenotypic correlation (𝑟 = 0.39 ± 0.04) and a high, between-line genetic correlation (𝑟 = 0.89 ± 0.03). A significant decrease of the genetic correlation was observed when comparing more different temperatures. Finally, among 7 other morphometrical traits which were measured on the same set of lines, 3 provided a significant positive genetic correlation with abdominal bristles: thoracic bristles, abdomen pigmentation and thoracic pigmentation.

    • A new measure to study phylogenetic relations in the brown algal order Ectocarpales: The ``codon impact parameter"

      Smarajit Das Jayprokas Chakrabarti Zhumur Ghosh Satyabrata Sahoo Bibekanand Mallick

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      We analyse forty-seven chloroplast genes of the large subunit of RuBisCO, from the algal order Ectocarpales, sourced from GenBank. Codon-usage weighted by the nucleotide base-bias defines our score called the codon-impact-parameter. This score is used to obtain phylogenetic relations amongst the 47 Ectocarpales. We compare our classification with the ones done earlier.

    • Structure and function of the spermathecal complex in the phlebotomine sandfly Phlebotomus papatasi Scopoli (Diptera: Psychodidae): I. Ultrastructure and histology

      K Ilango

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      Females of phlebotomine sandflies (Diptera: Psychodidae) possess highly variable spermathecae that present several important taxonomic characters. The cause of this diversity remains a neglected field of sandfly biology, but may possibly be due to female post-mating sexual selection. To understand this diversity, a detailed study of the structure and function of the spermathecal complex in at least one of the species was a prerequisite. Using scanning and transmission electron microscopy, described here is ultrastructure of the spermathecal complex in the sand fly, Phlebotomus papatasi Scopoli. The spermathecal complexes are paired; each consists of a long spermathecal duct, a cylindrical spermathecal body, and a spherical spermathecal gland. Muscle fibres, nerves, tracheoles, and vascular sinuses connect the spermathecal body and duct through the epithelial layers. Spermathecal gland is formed by a typical insect epidermis and consisting of an epithelial layer of class-1 epidermal cells and elaborate glandular cells of class-3 epidermal cells, each having both receiving and conducting ductules (i.e. “end apparatus”) and a “cytological apodeme”, which is a newly described cell structure. The spermathecal body and duct are lined by class-1 epidermal cells and a cuticle, and are enveloped by a super-contracting visceral muscular system. The cuticle consists of rubber-like resilin, and its fibrillar arrangement and chemical nature are described. A well-developed neuromuscular junction exists between the spermathecal gland and the spermathecal body, which are connected to each other by a nerve and a muscle. The spermathecal complexes of the sandfly are compared with those of other insect species. The physiological role and possible evolutionary significance of the different parts of spermathecal complex in the sandfly are inferred from the morphology and behaviour. Post-mating sexual selection may be responsible for the structural uniqueness of the spermathecal complex in phlebotomine sandflies.

    • Structure and function of the spermathecal complex in the phlebotomine sandfly Phlebotomus papatasi Scopoli (Diptera: Psychodidae): II. Post-copulatory histophysiological changes during the gonotrophic cycle

      K Ilango

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      The spermathecal complex of Phlebotomus papatasi Scopoli (Diptera: Psychodidae) undergoes histological and physiological changes during its gonotropic cycle. The present histochemical study revealed a mucopolysaccharide secretory mass in the spermathecae of the newly emerged sandfly. Sperm competition occurs when two or more males compete to fertilize an ovum in the female reproductive tract. In this study, spermatophores of two or more competing males were deposited at the base of the spermathecal ducts, which originate from the female bursa copulatrix. This suggests that females play a role in sperm displacement, which is defined as any situation in which the last male to mate with a female fertilizes maximum number her eggs. A blood meal ingested by the female for ovary development and egg laying stimulates the release of sperm from the spermatophore. The spermatozoa then migrate to the lumen of the spermatheca. The ultrastructure of spermatozoa comprises a head with double-layered acrosomal perforatorium, an elongate nucleus, and the axoneme with a 9 + 9 + 0 flagellar pattern. This axomene differs from the aflagellate axoneme of other Psychodinae. Morphological changes, such as the casting off of the acrosomal membrane, and histological changes in the spermatophore are also described. Mating plugs that have been described previously in sandflies appear to be artefacts. Females of P. papatasi may be inseminated more than once during each gonotrophic cycle, and additional inseminations may be necessary for each cycle. The relationships between the volumes of the sperm and the spermatheca were calculated to determine sperm utilization and fecundity of P. papatasi. As the females of P. papatasi mate polyandrously, the anatomical and physiological complexity of the spermathecal complex may be related to post-copulatory sexual selection.

    • Nutrients and toxin producing phytoplankton control algal blooms – a spatio-temporal study in a noisy environment

      Ram Rup Sarkar Horst Malchow

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      A phytoplankton-zooplankton prey-predator model has been investigated for temporal, spatial and spatio-temporal dissipative pattern formation in a deterministic and noisy environment, respectively. The overall carrying capacity for the phytoplankton population depends on the nutrient level. The role of nutrient concentrations and toxin producing phytoplankton for controlling the algal blooms has been discussed. The local analysis yields a number of stationary and/or oscillatory regimes and their combinations. Correspondingly interesting is the spatio-temporal behaviour, modelled by stochastic reaction-diffusion equations. The present study also reveals the fact that the rate of toxin production by toxin producing phytoplankton (TPP) plays an important role for controlling oscillations in the plankton system. We also observe that different mortality functions of zooplankton due to TPP have significant influence in controlling oscillations, coexistence, survival or extinction of the zooplankton population. External noise can enhance the survival and spread of zooplankton that would go extinct in the deterministic system due to a high rate of toxin production.

    • Sugar signalling and gene expression in relation to carbohydrate metabolism under abiotic stresses in plants

      Anil K Gupta Narinder Kaur

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      Sucrose is required for plant growth and development. The sugar status of plant cells is sensed by sensor proteins. The signal generated by signal transduction cascades, which could involve mitogen-activated protein kinases, protein phosphatases, Ca2+ and calmodulins, results in appropriate gene expression. A variety of genes are either induced or repressed depending upon the status of soluble sugars. Abiotic stresses to plants result in major alterations in sugar status and hence affect the expression of various genes by down- and up-regulating their expression. Hexokinase-dependent and hexokinase-independent pathways are involved in sugar sensing. Sucrose also acts as a signal molecule as it affects the activity of a proton-sucrose symporter. The sucrose transporter acts as a sucrose sensor and is involved in phloem loading. Fructokinase may represent an additional sensor that bypasses hexokinase phosphorylation especially when sucrose synthase is dominant. Mutants isolated on the basis of response of germination and seedling growth to sugars and reporter-based screening protocols are being used to study the response of altered sugar status on gene expression. Common cis-acting elements in sugar signalling pathways have been identified. Transgenic plants with elevated levels of sugars/sugar alcohols like fructans, raffinose series oligosaccharides, trehalose and mannitol are tolerant to different stresses but have usually impaired growth. Efforts need to be made to have transgenic plants in which abiotic stress responsive genes are expressed only at the time of adverse environmental conditions instead of being constitutively synthesized.

    • Subject Index

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    • Author Index

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    • Acknowledgements

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