Molecular Genetics

Substance misuse is influenced by multiple genetic and environmental factors. Recent research has identified a number of potential genetic markers of risk and those associated with drug reward substrates show particular promise. The current study reexamines the extant published data of the association between the D 2 dopamine receptor (DRD2) gene minor Taq 1A (A 1 ) allele and substance misuse risk. A series of meta-analyses was performed on 64 studies examining DRD2 A 1 + allelic status and substance misuse. In addition, personality was examined as a possible endophenotype. Significant association was found between the A 1 allele and severe substance dependence in both Caucasian and non-Caucasian groups. The data did not support a significant association between the A 1 allele and personality features. While the specific mechanism underlying these associations requires further elucidation, this genetic marker shows promise as a marker of brain reinforcement processes. Possible ways of utilising the A 1 allele to inform prevention and treatment initiatives are discussed. D

The mechanisms that drive normal B cell differentiation and activation are frequently subverted by B cell lymphomas for their unlimited growth and survival. B cells are particularly prone to malignant transformation because the machinery used for antibody diversification can cause chromosomal translocations and oncogenic mutations. The advent of functional and structural genomics has greatly accelerated our understanding of oncogenic mechanisms in lymphomagenesis. The signaling pathways that normal B cells utilize to sense antigens are frequently derailed in B cell malignancies, leading to constitutive activation of prosurvival pathways. These malignancies co-opt transcriptional regulatory systems that characterize their normal B cell counterparts and frequently alter epigenetic regulators of chromatin structure and gene expression. These mechanistic insights are ushering in an era of targeted therapies for these cancers based on the principles of pathogenesis.

High-risk screening Urine Liquid chromatography-tandem mass spectrometry LC-MS/MS MPS I MPS II, MPS VI Biomarker Mucopolysaccharidoses (MPSs) are complex storage disorders caused by specific lysosomal enzyme deficiencies, resulting in the accumulation of glycosaminoglycans (GAGs) in urine, plasma, as well as in various tissues. We devised and validated a straightforward, but accurate and precise tandem mass spectrometry methodology coupled to high performance liquid chromatography (LC-MS/MS) for the quantification of GAGs in urine. The method is applicable to the investigation of patients with MPS I, II, and VI, by quantifying dermatan sulfate (DS) and heparan sulfate (HS) in urine. We analyzed urine samples from 28 MPS patients, aged 1 to 42 years, and 55 control subjects (41 days to 18 years old). Levels of DS and HS in urine from healthy controls of all ages were below the limit of quantification. The levels of DS and HS in urine from 6 treated patients with MPS I were lower than in 6 untreated patients in DS (0.7-45 vs 9.3-177 mg/mmol creat) and HS (0-123 mg/mmol creatinine vs 38-418 mg/mmol creatinine); similar results were obtained for 9 patients with MPS II and 7 patients with MPS VI. Analyses were performed on as little as 250 μL of urine. Methanolysis took 75 min per sample; the total analysis run time for each LC-MS/MS injection was 8 min. Results indicate that the method is applicable to a wide variety of situations in which high accuracy and precision are required, including the evaluation of the effectiveness of existing and emerging treatments.

Diego Wyszynski has done an admirable job of assembling a collection of reviews of various aspects of the etiology and treatment of oral clefts. This 500-page monograph is divided into three sections: "Basic Principles" (embryology, clinical features, epidemiology, and genetics); "Treatment" (surgery, speech care, dentistry, otolaryngology, etc.); and "Public Health Issues" (prevention, insurance coverage, ethical issues, and education). As with all edited monographs, some essays are more scholarly than others; a few are rather poor. Nevertheless, this book achieves its stated intention of "addressing issues that are relevant to clinicians, researchers, and family members." Of particular value to clinicians and well-informed family members is the section on treatment. The treatment of patients with oral clefts, from birth to adulthood, requires a well-coordinated team effort. This section does a good job of conveying the global nature and specific details of this comprehensive care. Thus, chapters 24-34 are highly recommended, as are chapters 36-39 in the section on public health issues. One important misstep in this presentation is the lack of succinct clarity regarding our present knowledge of oral cleft etiology. In 1949, Curt Stern, writing in his text Principles of Human Genetics, noted: "Harelip and cleft palate are developmental abnormalities which have a genetic basis. In many pedigrees, they depend on the cooperation of specific alleles at several autosomal loci, and, in addition, require the presence of mostly uncontrolled environmental factors." In 2002, there is little else we can add. F. Clarke Fraser, in his foreword to Wyszynski's monograph, cogently tells us why: "Epidemiologic studies are hampered by the fact that, for an uncommon multifactorial trait, it is virtually impossible to assemble groups large enough to allow statistically valid comparisons of potentially significant factors such as prenatal events, susceptibility genes, social class, and race." Unfortunately, one is left to conclude that much of what is contained in chapter 3 and chapters 16-22 is much ado about very little. I hope that, someday, this will be different. Today, at bottom, we must rely entirely on the information in Carmella Stadter's chapter (31) on genetic counseling and empiric risk calculation. This message is lost amid the genomic hype. In a book "addressing issues … relevant to clinicians … and family members," this should have been in neon. On balance, this monograph has more to recommend it than not. It should be on the shelf of all those with a need to know about clefts.

Background: The bacterial luciferase (lux) gene cassette consists of five genes (luxCDABE) whose protein products synergistically generate bioluminescent light signals exclusive of supplementary substrate additions or exogenous manipulations. Historically expressible only in prokaryotes, the lux operon was re-synthesized through a process of multibicistronic, codon-optimization to demonstrate for the first time self-directed bioluminescence emission in a mammalian HEK293 cell line in vitro and in vivo. Methodology/Principal Findings: Autonomous in vitro light production was shown to be 12-fold greater than the observable background associated with untransfected control cells. The availability of reduced riboflavin phosphate (FMNH 2) was identified as the limiting bioluminescence substrate in the mammalian cell environment even after the addition of a constitutively expressed flavin reductase gene (frp) from Vibrio harveyi. FMNH 2 supplementation led to a 151fold increase in bioluminescence in cells expressing mammalian codon-optimized luxCDE and frp genes. When injected subcutaneously into nude mice, in vivo optical imaging permitted near instantaneous light detection that persisted independently for the 60 min length of the assay with negligible background. Conclusions/Significance: The speed, longevity, and self-sufficiency of lux expression in the mammalian cellular environment provides a viable and powerful alternative for real-time target visualization not currently offered by existing bioluminescent and fluorescent imaging technologies.

Terpenes and sterols are complex molecules synthesized by equally complex biosynthetic pathways. Recent progress in using the tools of genetics, molecular genetics and genetic engineering to dissect triterpene metabolism in the cytosol, and terpene metabolism in the ...

Cystic fibrosis is the most common autosomal recessive potentially lethal hereditary disease. The disease is caused by mutations in the CFTR gene of the transmembrane peptide. CFTR, functioning as a cAMP-dependent chlorine channel, regulates chlorine and sodium channels, participates in water and ATP. As a result of a genetic defect, the secretion of the glands of external secretion thickens, its evacuation becomes more difficult, secondary changes occur in the organs.

This study evaluated the mutagenicity and antimutagenicity of inulin in a chromosomal aberration assay in cultures of the meristematic cells of Allium cepa. The treatments evaluated were as follows: negative control-seed germination in distilled water; positive control-aqueous solution of methyl methanesulfonate (10 μg/mL MMS); mutagenicity-aqueous solutions of inulin (0.015, 0.15, and 1.50 μg/mL); and antimutagenicity-associations between MMS and the different inulin concentrations. The antimutagenicity protocols established were pretreatment, simultaneous simple, simultaneous with pre-incubation, and post-treatment. The damage reduction percentage (DR%) was 43.33, 27.77, and 55.92% for the pre-treatment;-31.11, 18.51, and 7.03% for the simultaneous simple; 30.43, 19.12, and 21.11% for the simultaneous with pre-incubation; and 64.07, 42.96, and 53.70% for the post-treatment. The results indicated that the most effective treatment for inhibiting damages caused by MMS was the post-treatment, which was followed by the pretreatment, suggesting activity by bioantimutagenesis and desmutagenesis. The Allium cepa assay was demonstrated to be a good screening test for this type of activity because it is easy to perform, has a low cost, and shows DR% that is comparable to that reported studies that evaluated the prevention of DNA damage in mammals by inulin.

This work examines the cellular localization of holocarboxylase synthetase (HCS) and its association to chromatin during different stages of development of Drosophila melanogaster. While HCS is well known for its role in the attachment of biotin to biotin-dependent carboxylase, it also regulates the transcription of HCS and carboxylases genes by triggering a cGMP-dependent signal transduction cascade. Further, its presence in the nucleus of cells suggests additional regulatory roles, but the mechanism involved has remained elusive. In this study, we show in D. melanogaster that HCS migrates to the nucleus at the gastrulation stage. In polytene chromosomes, it is associated to heterochromatin bands where it co-localizes with histone 3 trimethylated at lysine 9 (H3K9met3) but not with the euchromatin mark histone 3 acetylated at lysine 9 (H3K9ac). Further, we demonstrate the association of HCS with the hsp70 promoter by immunofluorescence and chromatin immuno-precipitation (ChIP) of associated DNA sequences. We demonstrate the occupancy of HCS to the core promoter region of the transcriptionally inactive hsp70 gene. On heat-shock activation of the hsp70 promoter, HCS is displaced and the promoter region becomes enriched with the TFIIH subunits XPD and XPB and elongating RNA pol II, the latter also demonstrated using ChIP assays. We suggest that HCS may have a role in the repression of gene expression through a mechanism involving its trafficking to the nucleus and interaction with heterochromatic sites coincident with H3K9met3.

Plant seeds that have high phytate content are used as animal feed. Phytases, enzymes that catalyze the breakdown of phytate into inorganic phosphorus and myoinositol phosphate derivatives, have been intensively studied in recent years and gained immense attention because of their application in reducing phytate content in animal feed and food for human consumption, thus indirectly lowering environmental pollution caused by undigested phytate. This review is focused on summarising the current knowledge on recent developments of fungal and yeast phytases. Comparative account on diverse sources and physiological roles, molecular characteristics and regulation mechanisms of phytases are discussed. Phylogenetic relationship of phytases from different classes of fungi is studied in details. It is inferred on the basis of phylogeny that phytases from Ascomycetes and Basidiomycetes differ in the amino acid sequences, therefore they fall in separate clade in the tree. The prospective biotec...

Recent enhancement of the pool of known molecular markers for avocado has allowed the construction of the first moderately dense genetic map for this species. Over 300 SSR markers have been characterized and 163 of these were used to construct a map from the reciprocal cross of two Florida cultivars 'Simmonds' and 'Tonnage'. One hundred thirty-five primer pairs amplified 163 usable loci with 20 primer pairs amplifying more than one locus. 'Tonnage' was heterozygous for 152 (93%) loci, whereas 'Simmonds' was heterozygous for 64 (39%). Null alleles were identified at several loci. Linkage maps were produced for both reciprocal crosses and combined to generate a composite linkage map for the F1 population of 715 individuals. The composite map contains 12 linkage groups. Linkage groups ranged in size from 157.3 cM (LG2) to 2.4 cM (LG12) and the number of loci mapped per group ranged from 29 (LG1) to two (LG12). The total map length was 1,087.4 cM. Only seven markers were observed to have segregation distortion (α ≤ 0.05) across both sub-composite (reciprocal) maps. Phenotypic data from traits of horticultural interest are currently being collected on this population with the ultimate goal of identifying useful quantitative trait loci and the development of a marker-assisted selection program.

Advances in molecular genetics have opened opportunities to enhance strategies for genetic improvement of pigs by directly selecting on genes or chromosomal regions that harbor genes that affect traits of interest. In this paper, we review molecular technologies that have become available, the current state of the use of gene- or marker tests in pig breeding programs, and future prospects.

A molecular approach, based on the detection of ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) large subunit genes, was applied to investigate the distribution and diversity of autotrophic bacteria in groundwater systems. DNA extracts from 48 sampling stations, including a variety of pristine and polluted, shallow and deep-subsurface groundwater samples obtained from Germany and Austria, served as a template for the PCR amplification of form I (cbbL) and form II (cbbM) large subunit RubisCO genes. The majority of the samples (480%) contained two different forms of RubisCO. In 17 samples, all three forms of RubisCO were identified. PCR products from four selected groundwater habitats containing all three forms of RubisCO were used to construct clone libraries. Based on restriction fragment length polymorphism (RFLP) analysis, 109 RubisCO-clone-inserts were subjected to sequencing and phylogenetic analysis. With the exception of a form IA RubisCO sequence cluster obtained from deep subsurface samples, which was identical to the RubisCO genes described for Ralstonia metallidurans CH34, most sequences were distantly related to a variety of RubisCO species in chemolithoautotrophic Proteobacteria. Several sequences occurred in isolated lineages. These findings suggest that autotrophic bacteria with the capability to assimilate CO 2 via the Calvin Cycle pathway are widespread inhabitants of groundwater systems.

Cancer develops secondary to genetic and epigenetic changes of multiple genes.  The overall 5 year survival rate of cancer is less than 50%. The main reason of this poor outcome lies in the technical limitations of the methods used for the early detection of cancer. Cancer is characterized by extreme variation. ie.;  each cancer is different in its genetic profile.  It is possible to detect cancer cells, free “cancer DNA” or “cancer protein” from body fluid(blood, stool and sputum) of cancer patients even when the tumor mass is too small to be visible by radiologic study or endoscopy.  The objective of this lecture is to review existing new  genomic technology which aims to  detect cancer in its early stages by the testing of body fluids for a combination of multiple, cancer-related key genes and marker proteins.  The multiplex molecular test system for the early detection of  cancer, recognizes multiple cancer related genes and proteins, with high sensitivity and precision. It can also be applied to the management, treatment  and prevention of cancer by offering unique information on the nature of the genetic changes and alterations specifically associated with this particular cancer.

Three hundred and sixty-eight strains of aerobic, endospore-forming bacteria which included type and reference cultures of Bacillus and environmental isolates were studied. Overall similarities of these strains for 118 unit characters were determined by the SSM, S, and D p coefficients and clustering achieved using the UPGMA algorithm. Test error was within acceptable limits. Six cluster-groups were defined at 70% SSM, which corresponded to 69% Sp and 48-57% SJ. Groupings obtained with the three coefficients were generally similar but there were some changes in the definition and membership of cluster-groups and clusters, particularly with the SJ coefficient.

Tumbleweeds (Salsola species, section Kali) are road side and rangeland pest plants throughout the 48 contiguous states in the US. Three described tumbleweed species and two undescribed Salsola taxa occur in California. The known species are Russian thistle, Salsola tragus, introduced from Eurasia in the 1800s, Russian barbwire thistle, S. paulsenii, which grows in the desert regions of California, and is also native to Eurasia, and the recently identified S. kali subspecies austroafricana, possibly native to South Africa. Our goals were to investigate karyology, genome size, and molecular genetic affinities of the described species and the other taxa within their ranges in California using recently developed microsatellite loci, dominant nuclear DNA markers (RAPD and ISSR), and DNA sequence data. Chromosome counts and genome size assessments made with flow cytometry were compared. These analyses indicated that one undescribed taxon is a new allopolyploid hybrid between S. tragus and S. kali subspecies austroafricana, and the other undescribed taxon appears to be a complex hybrid involving all three described species. The invasion potentials for the hybrid taxa are unknown. Tumbleweeds are the focus of biological controls efforts but the identification of suitable agents for the hybrid taxa may be problematic because of the large amount of genetic variability encompassed within this evolving Salsola complex.

The present review summarizes the current state of knowledge about the genetics of pain-related phenomena and illustrates the scope and power of genetic approaches to the study of pain. We focus on work performed in our laboratories in Jastrzebiec, Poland; Portland, OR; and Los Angeles, which we feel demonstrates the continuing usefulness of classical genetic approaches, especially when used in combination with newly available molecular genetic techniques.

In 2003, an international working group last reported on recommendations for diagnosis, response assessment, and treatment outcomes in acute myeloid leukemia (AML). Since that time, considerable progress has been made in elucidating the molecular pathogenesis of the disease that has resulted in the identification of new diagnostic and prognostic markers. Furthermore, therapies are now being developed that target disease-associated molecular defects. Recent developments prompted an international expert panel to provide updated evidence- and expert opinion–based recommendations for the diagnosis and management of AML, that contain both minimal requirements for general practice as well as standards for clinical trials. A new standardized reporting system for correlation of cytogenetic and molecular genetic data with clinical data is proposed.

Rapid and prolific shoot regeneration via direct organogenesis was induced from leaf explants of Patchouli, Pogostemon cablin (Blanco) Benth., an aromatic plant of immense industrial value, on Murashige and Skoog (MS) medium with benzylaminopurine (BAP) and ␣-naphthaleneacetic acid (NAA) within 4 weeks. The adventitious shoot bud induction and plant regeneration were greatly influenced by the origin, age of donor plant, and leaf position on stem. Leaf explants prepared from in vivo plants of different ages showed higher regeneration response as compared to the explants from in vitro plantlets of respective age. The shoot regeneration ability of explants was significantly related to the age of the donor plants as well as the leaf position on the stem. The highest number of shoots (94.6/explants) was obtained from 96.2% of leaf explants derived from the leaves located on the second node of the 3-monthold in vivo plants on Murashige and Skoog's (MS) medium containing 2.5 M benzylaminopurine (BAP) and 0.5 M naphthaleneacetic acid (NAA). Incorporation of 1.0 M gibberellic acid (GA 3) in MS medium significantly improved the shoot elongation (1.8-fold) within 2 weeks in 95% of the cultures. Regenerated shoots rooted spontaneously on growth regulator-free half strength MS medium and were successfully hardened and transferred to nursery with 96-100% survival rate. Genetic fidelity of the in vitro derived plants was assessed using random amplified polymorphic DNA (RAPD). Fourteen arbitrary decamers displayed same banding profile within all the micropropagated plants and in vivo explant donor plant. The molecular analysis complemented and compared well and showed genetic stability in the plants regenerated through direct shoot bud differentiation from leaf explants. The gas chromatogram of the extracted oils from in vitro derived plants and the mother stock plant showed similar essential oil profiles. Rapid and high multiplication frequency, molecular, genetic and essential oil content stability ensure the efficacy of the protocol developed for the production of this industrially important aromatic plant.

Cryptic species of the digeneans, Cercaria batillariae (Heterophyidae) and an undescribed philophthalmid, were detected using polymerase chain reaction-based restriction fragment-length polymorphism methodology and sequence analyses. These digeneans were all collected from the same species of gastropod first intermediate host, Batillaria cumingi (ZBatillaria attramentaria). The mitochondrial cytochrome oxidase subunit 1 gene (approximately 800 bp) and nuclear internal transcribed spacer 1 gene (approximately 400 bp) were used for species level discrimination. Restriction fragment-length polymorphism analyses of cytochrome oxidase subunit 1 gene showed that C. batillariae included 10 distinguishable fragment patterns, and the philophthalmid included five patterns. On the basis of subsequent sequence analyses, the restriction fragment length polymorphism patterns of C. batillariae were grouped into eight phylogenetically distinct lineages and those of the philophthalmid into three phylogenetically distinct lineages. There was no evidence of gene flow among the different lineages due to the lack of heterozygosity within the observed internal transcribed spacer 1 gene fragment patterns. This suggests that all of these lineages are different species. Most of these species were widespread, but some exhibited restricted geographic distributions. We discuss the implications of these findings for host specificity of these trematodes. These results demonstrate the utility of genetic analysis to distinguish species of morphologically similar trematodes. Hence, trematode species diversity may often be underestimated when species identifications are limited to morphological features. q

and Introduction Abstract In the choice betvveen male or developrnent !s genetically determined. either testis ar ()vary, This by genes that hm',jrl been disc;()Vefed knockout experiments in mice. The pathvvay is better knovvn thi:'m the s cascade to testicular differentiation. in thi~cascade: MolO

Substrate reduction therapy (SRT) is considered to be a potential therapeutic option for juvenile GM2 gangliosidosis (jGM2g). We evaluated the efficacy of SRT in jGM2g, assessing neurological, neuropsychological and brain magnetic resonance imaging (MRI) outcomes over a 24-month period of treatment. In an open-label and single-center study, five jGM2g patients (mean age 14.6+/-4.5 years) received oral miglustat at doses of 100-200mg t.i.d. adjusted to body surface area. Patients underwent general and neurological examinations, neuropsychological, electrophysiological, and brain MRI studies. All patients showed neurological deterioration over the period of the study, with particularly notable worsening of gait, speech and coordination. One patient experienced acute psychosis, and another showed worsening of pre-existing epilepsy. Some neuropsychological tests showed no evidence of deterioration in the three patients with high enough cognitive functioning for reliable assessment. Profound cognitive impairment in two children precluded neuropsychological evaluation. In four patients, evaluation of brain MRI showed no changes in white matter signal abnormalities and cerebellar atrophy noted at baseline, while one patient showed progression of cerebellar and supratentorial brain atrophy. Transmission electron microscopy analysis of peripheral mononuclear cells showed reduction of intracytoplasmatic inclusions with treatment. SRT with miglustat of patients with jGM2g failed to ameliorate progressive neurological deterioration, but apparently no worsening of some areas of cognitive function tested and brain MRI lesions was noted over 24 months of treatment. The results must be interpreted with care owing to the small sample of patients and the lack of a control-arm.

In atomic force microscopy, the tip experiences electrostatic, van der Waals, and hydration forces when imaging in electrolyte solution above a charged surface. To study the electrostatic interaction force vs distance, curves were recorded at different salt concentrations and pH values. This was done with tips bearing surface charges of different sign and magnitude (silicon nitride, A1203, glass, and diamond) on negatively charged surfaces (mica and glass). In addition to the van der Waals attraction, neutral and negatively charged tips experienced a repulsive force. This repulsive force depended on the salt concentration. It decayed exponentially with distance having a decay length similar to the Debye length. Typical forces were about 0.1 nN strong. With positively charged tips, purely attractive forces were observed. Comparing these results with calculations showed the electrostatic origin of this force.

PROEFSCHRIFT Ter verkrijging van de graad van doctor aan de Erasmus Universiteit Rotterdam op gezag van de rector magnificus Prof. Dr. P.W.c. Akkermans M.A. en volgens besluit van het College voor Promoties. De openbare verdediging zal plaatsvinden op woensdag 20 december 1995 om 9.45 uur door WILLEM VERMEULEN geboren te Hardinxveld·Giessendam PROMOTIECOMMISSIE Pomotoren:

Urogenital birth defects are one of the common phenotypes observed in hereditary human disorders. In particular, limb malformations are often associated with urogenital developmental abnormalities, as the case for Hand-foot-genital syndrome displaying similar hypoplasia/agenesis of limbs and external genitalia. Split-hand/split-foot malformation (SHFM) is a syndromic limb disorder affecting the central rays of the autopod with median clefts of the hands and feet, missing central fingers and often fusion of the remaining ones. SHFM type 1 (SHFM1) is linked to genomic deletions or rearrangements, which includes the distal-less-related homeogenes DLX5 and DLX6 as well as DSS1. SHFM type 4 (SHFM4) is associated with mutations in p63, which encodes a p53-related transcription factor. To understand that SHFM is associated with urogenital birth defects, we performed gene expression analysis and gene knockout mouse model analyses. We show here that Dlx5, Dlx6, p63 and Bmp7, one of the p63 downstream candidate genes, are all expressed in the developing urethral plate (UP) and that targeted inactivation of these genes in the mouse results in UP defects leading to abnormal urethra formation. These results suggested that different set of transcription factors and growth factor genes play similar developmental functions during embryonic urethra formation. Human SHFM syndromes display multiple phenotypes with variations in addition to split hand foot limb phenotype. These results suggest that different genes associated with human SHFM could also be involved in the aetiogenesis of hypospadias pointing toward a common molecular origin of these congenital malformations.

When Princess Augusta and Lord Bute, followed by Sir Joseph Banks and King George III, started gathering plants at Kew, conservation on the site can be said to have begun. Although the primary motive then was to assist the expansion of the British Empire and trade, rare plants were gathered and some became rare or extinct in the wild as their habitats were destroyed. The primary motive in the nineteenth century was not conservation, but the history of conservation at the Royal Gardens at Kew dates back to its very origins. Subsequent regimes at Kew maintained and added to the collections thereby adding to their conservation value. Many early collections are of species now listed within the IUCN categories of endangerment. Environmental awareness and concern had begun by the time that Professor Jack Heslop-Harrison became director and he was the first director actively to initiate specific conservation programmes such as seed banking and work on red data books. From then on conservation became an integral part of the work programme of Kew and the focus on conservation has increased with each subsequent director. This eventually led to the transformation of the embryonic seed banking activities into the Millennium Seed Bank, the largest and most important bank in the world for the conservation of the seeds of wild species. It currently holds just over ten percent of all seed plant species. Conservation at Kew over the past three decades has very much been a balance between ex situ work and in situ activities to help conservation in the overseas areas where Kew scientists have experience. Throughout the history of the gardens there has been a vital interest in economic botany that has developed from moving plants around the empire to much work on the sustainable use of plants and ecosystems thereby better equipping the institution to subsequently work on in situ conservation. Significant conservation activity at Kew has been possible because it is being supported by a solid research programme that includes such areas as systematics and molecular genetics and laboratories, a large herbarium and a large library. Kew has played an important role in stimulating conservation work elsewhere and such units as the Threatened Plants Unit of IUCN and Botanic Gardens Conservation International (BGCI) have their roots in Kew. Among other important conservation initiatives have been the creation of a unit to work with the implementation of the CITES treaty on the trade of endangered plants and a legal unit to work on issues of the Convention on Biological Diversity (CBD). There is no doubt that the Royal Botanic Gardens at Kew is at the forefront of plant conservation.

Eight isolates of Sclerotium rolfsii from four strategically geographical sites of Bangladesh were characterized and their cultural properties like average linear mycelial growth, colony colour, colony consistency, growth pattern and sclerotia formation were studied. Isolates varied in mycelial growth and other growth characteristics and were grouped into three. The highest linear growth was displayed by S8. DNA concentration of eight isolates varied from 1150-7200 ng/µl. DNA fingerprinting by RAPD prompted the grouping of isolates. Selected 3 primers generated 20 bands with size ranging from 100-1500 bp. Out of the 20 bands, 9 bands (45%) were polymorphic and 11 bands (55%) were monomorphic among the eight isolates of Sclerotium rolfsii. The coefficient of gene differentiation (Gst) was 1.000 reflecting the existence of high level of genetic variations among the 8 isolates. The lowest genetic distance and highest inter isolate similarity was found in S1 and S2 which would be homogeneous. The highest genetic distance and lowest inter isolate similarity found in S3, S7 and S3, S8 pair which would be most divergent isolates. The cluster analysis also revealed that S3, S7 and S8 belong to different clusters. All five varieties of eggplant and tomatoes were graded as susceptible when inoculated with eight isolates. Plant mortality 93.33% was recorded in S4, S6 and in S8. Considering the isolate factor the most virulent isolate would be S8 whereas the less virulent isolate would be S2 and S7. Host plant of S8 was tomato collected from Thakurgaon. S2 and S7 were collected from BAU farm and Dinajpur and host plants were lentil and tomato respectively. It is evident that Sclerotium rolfsii from Thakurgaon on host tomato is more virulent.

A joint evaluation of all animals, uses all additive genetic relationships uses all data on all animal jointly It works as a linear model (correcting different effects for each other), jointly estimates animal effects and fixed effects (herds) but has Selection Index properties (Regression with " heritability ")  1 0 0 0 1 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0 1 0 0 0 0 1

New insights into the phenomenon of systemic acquired resistance have been gained in recent years, by the use of techniques in molecular genetics and biology that have replaced the largely descriptive approach of earlier work. The isolation of mutants in the signal transduction pathway from induction to expression of resistance as well as the use of transgenic plants over-expressing or suppressing the expression of putative candidate genes involved in systemic acquired resistance and its signalling have identified several steps in the establishment of plant resistance. In this review the latest developments implicating salicylic acid as a signal molecule in systemic resistance are discussed and contrasted with new signalling pathways which, seemingly, are based on alternative mechanisms.

The genus Podocnemis comprises six living species, including P. erythrocephala (irapuca -red-headed river turtle). Data are available concerning the reproductive biology of the species of the genus Podocnemis, but little is known about their reproductive strategies. Considering the total lack of such data for P. erythrocephala, and with the goal of contributing information on their mode of reproduction, we examined the relationships among individuals of nests of this turtle, using microsatellite markers. Using four microsatellite loci, we analyzed the progeny in six nests from two localities in the Brazilian Amazon (Santa Isabel do rio Negro and Parintins). All juveniles from each nest were analyzed. The genotypes of each juvenile from each nest were identified, and because a sample of female DNA was not available, the maternal genotype was inferred from homozygous individuals in each nest. We found that this species is promiscuous; there was multiple paternity in five of the six nests analyzed. In addition to being important for the understanding of evolutionary and genetic processes, this type of information will be useful for chelonian management projects. Our data suggest one possible ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 9 (1): 435-440 (2010) C. Fantin et al. difference between reproductive patterns of the different populations. This multi-paternal condition may be a natural reproductive strategy for the preservation of the genetic diversity of this species.

Introduction Mowat–Wilson syndrome is a congenital syndrome caused by a defect of the transcriptional repressor ZFHX1B (SIP1) gene on the chromosome 2q22–q23. The genotype–phenotype analysis confirmed that ZFHX1B deletions and mutations result in a recognizable facial dysmorphism with a multiple congenital anomaly and mental retardation. Case report This report is about one new patient from Croatia with the typical phenotype. Molecular genetic studies showed the novel mutation in ZFHX1B (exon 8: c.2372del C; p.T791fsX816). This mutation has not been reported before. The literature is reviewed. Conclusion Mowat–Wilson syndrome is a newly described congenital syndrome and should be considered in any individual with characteristic facial features and mental retardation in associations with congenital malformations.

KEYWORDS Recurrent pregnancy loss; immunity; pro-inflammatory cytokines; homogenous samples ... ABSTRACT It is a well-known fact that pro-inflammatory cytokines exert an adverse effect on conceptus and result in pregnancy failure and there are mutations reported in ...

Currently, molecular diagnosis of haemophilia A and B (HA and HB) highlights the excess risk-inhibitor development associated with specific mutations, and enables carrier testing of female relatives and prenatal or preimplantation genetic diagnosis. Molecular testing for HA also helps distinguish it from von Willebrand disease (VWD). Next-generation sequencing (NGS) allows simultaneous investigation of several complete genes, even though they may span very extensive regions. This study aimed to evaluate the usefulness of a molecular algorithm employing an NGS approach for sequencing the complete F8, F9 and VWF genes. The proposed algorithm includes the detection of inversions of introns 1 and 22, an NGS custom panel (the entire F8, F9 and VWF genes), and multiplex ligation-dependent probe amplification (MLPA) analysis. A total of 102 samples (97 FVIII- and FIX-deficient patients, and five female carriers) were studied. IVS-22 screening identified 11 out of 20 severe HA patients and ...

Background: The Elaeis oleifera genetic materials were assembled from its center of diversity in South and Central America. These materials are currently being preserved in Malaysia as ex situ living collections. Maintaining such collections is expensive and requires sizable land. Information on the genetic diversity of these collections can help achieve efficient conservation via maintenance of core collection. For this purpose, we have applied fourteen unlinked microsatellite markers to evaluate 532 E. oleifera palms representing 19 populations distributed across Honduras, Costa Rica, Panama and Colombia. Results: In general, the genetic diversity decreased from Costa Rica towards the north (Honduras) and southeast (Colombia). Principle coordinate analysis (PCoA) showed a single cluster indicating low divergence among palms. The phylogenetic tree and STRUCTURE analysis revealed clusters based on country of origin, indicating considerable gene flow among populations within countries. Based on the values of the genetic diversity parameters, some genetically diverse populations could be identified. Further, a total of 34 individual palms that collectively captured maximum allelic diversity with reduced redundancy were also identified. High pairwise genetic differentiation (Fst > 0.250) among populations was evident, particularly between the Colombian populations and those from Honduras, Panama and Costa Rica. Crossing selected palms from highly differentiated populations could generate off-springs that retain more genetic diversity. Conclusion: The results attained are useful for selecting palms and populations for core collection. The selected materials can also be included into crossing scheme to generate offsprings that capture greater genetic diversity for selection gain in the future.

Human endogenous retroviruses (HERVs), which constitute a significant part of the human genome, might have a serious impact on primate evolution. Over a hundred insertions of HERV-K(HML-2) family members distinguish the human genome from other primate genomes. However, only three cases of insertion polymorphisms have been reported so far, all for endogenous HERV-K proviruses. This suggests that some retroviral integrations occurred rather recently in human genome evolution. In this report, we describe a very rare case of true insertion polymorphism of a solitary HERV-K LTR in the human genome. Distribution of the LTR-containing allele was tested in 5 Africans and 83 individuals from three Russian populations. The allele frequency appeared to be relatively high in populations of both European and Asian origin. The detected polymorphic LTR could be a useful molecular genetic marker of the corresponding genomic region.

Plant breeders have used disease resistance genes (R genes) to control plant disease since the turn of the century. Molecular cloning of R genes that enable plants to resist a diverse range of pathogens has revealed that the proteins encoded by these genes have several features in common. These findings suggest that plants may have evolved common signal transduction mechanisms for the expression of resistance to a wide range of unrelated pathogens. Characterization of the molecular signals involved in pathogen recognition and of the molecular events that specify the expression of resistance may lead to novel strategies for plant disease control.

The fungus Mycosphaerella fijiensis is the causative agent of black sigatoka, which is one of the most destructive diseases of banana plants. Infection with this pathogen results in underdeveloped fruit, with no commercial value. We analyzed the distribution of the M. fijiensis mating-type system and its genetic variability using M13 phage DNA markers. We found a 1:1 distribution of mating-type alleles, indicating MAT1-1 and MAT1-2 idiomorphs. A polymorphism analysis using three different primers for M13 markers showed that only the M13 minisatellite primers generated polymorphic products. We then utilized this polymorphism to characterize 40 isolates from various Brazilian states. The largest genetic distances were found between isolates from the ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 12 (1): 443-452 (2013) C.B. Queiroz et al. same location and between isolates from different parts of the country. Therefore, there was no correlation between the genetic similarity and the geographic origin of the isolates. The M13 marker was used to generate genetic fingerprints for five isolates; these fingerprints were compared with the band profiles obtained from inter-simple sequence repeat (UBC861) and inter-retrotransposon amplified polymorphism analyses. We found that the M13 marker was more effective than the other two markers for differentiating these isolates.

Рассмотрены концептуальные положения молекулярной биологии, важнейшие признаки формирования индивидуальных признаков живого организма, молекулярные процессы, лежащие в основе функционирования нервной системы человека, его психики,а также методы современной психогенетики и их возможности.
Приведены данные о влиянии конкретных генов на психологические признаки животных и человека и молекулярных механизмах такого влияния. Освещены вопросы истории возникновения психогенетики, перспективы ее развития, современного представления об эволюции человеческой психики и этические аспекты психогенетики. Для студентов учреждений высшего профессионального образования.