Molecular genetics in clinical practice: evolution of a DNA diagnostic service.

The development of a molecular genetics diagnostic service over a three year period was studied in a National Health Service region with a population of three million. Starting from a time when few diagnostic applications were possible, the number of disorders and the overall demand had grown rapidly. Conditions for which molecular genetic diagnosis had been provided included Duchenne and Becker muscular dystrophy, myotonic dystrophy, Huntington''s disease, and cystic fibrosis. Of 405 requests for diagnosis, 151 (37%) related to determination of carrier state, 187 (46%) to determining the feasibility of future prenatal diagnosis, and 67 (17%) were prenatal diagnostic biopsy samples, almost exclusively of first trimester chorion. DNA samples for future diagnostic use with a wide range of genetic disorders had also been banked. The study showed a need for close integration with clinical genetics services to allow satisfactory genetic counselling and interpretation of risks.     

Induction and release of secondary seed dormancy in genetically pure lines of Avena fatua

Induction and release of secondary dormancy in genetically pure dormant (AN-51, Mont 73) and non-dormant (CS-40, SH-430) lines of wild oat ( Avena fatua L.) were studied. These lines differed with regard to the optimal period of anaerobiosis necessary for induction of dormancy, and/or the degree (% of seeds acquiring dormancy) and duration of the dormancy induced. Secondary dormancy could be induced more effectively in the after-ripened seeds of dormant lines than in the non-dormant lines, where only a short-term dormancy could be induced (in 5–7 week-old-seeds). Higher anaerobiosis temperatures were more effective in inducing dormancy in all lines studied. Thus, as with primary dormancy, wild oat biotypes exhibit genetic variability in their secondary dormancy behaviour and factors like temperature can modify the expression of this trait.
The germination stimulants kinetin, isopentenyl adenine, sodium azide, potassium nitrate, ethanol and substituted phthalimides, which break primary dormancy in wild oats, stimulated germination of secondarily dormant seeds (line AN-51). Since these chemicals are structurally diverse, primary and secondary dormancies appear to be similar in part in their regulation.
Salicylhydroxamic acid, an inhibitor of cyanide-insensitive (alternative) respiration, did not inhibit: 1, spontaneous release of secondary dormancy in the line SH-430; and 2, stimulation of germination of secondarily dormant AN-51 seeds by various chemicals (except azide), suggesting that this respiratory pathway is not necessary for the release of induced dormancy.     

Effect of soil solarization on nodulation,infection by mycorrhizal fungi and yield of cowpea

Solarization of soil was found beneficial for plant growth in cowpea under field conditions. Root nodulation, infection by mycorrhizal fungi and yield were higher in plants grown in solarized soil. These increases were to the extent of 104.7, 20.0 and 23.7 per cent respectively when compared to control treatment without solarization.     

Purification of Mitochondrial Glutamate Dehydrogenase from Dark-Grown Soybean Seedlings

Proteins in extracts from cotyledons, hypocotyls, and roots of 5-d-old, dark-grown soybean (Glycine max L. Merr. cv Williams) seedlings were separated by polyacrylamide gel electrophoresis. Three isoforms of glutamate dehydrogenase (GDH) were resolved and visualized in gels stained for GDH activity. Two isoforms with high electrophoretic mobility, GDH1 and GDH2, were in protein extracts from cotyledons and a third isoform with the lowest electrophoretic mobility, GDH3, was identified in protein extracts from root and hypocotyls. Subcellular fractionation of dark-grown soybean tissues demonstrated that GDH3 was associated with intact mitochondria. GDH3 was purified to homogeneity, as determined by native and sodium dodecyl sulfate-polyacrylamide gels. The isoenzyme was composed of a single 42-kD subunit. The pH optima for the reductive amination and the oxidative deamination reactions were 8.0 and 9.3, respectively. At any given pH, GDH activity was 12- to 50-fold higher in the direction of reductive amination than in the direction of the oxidative deamination reaction. GDH3 had a cofactor preference for NAD(H) over NADP(H). The apparent Michaelis constant values for [alpha]-ketoglutarate, ammonium, and NADH at pH 8.0 were 3.6, 35.5, and 0.07 mM, respectively. The apparent Michaelis constant values for glutamate and NAD were 15.8 and 0.10 mM at pH 9.3, respectively. To our knowledge, this is the first biochemical and physical characterization of a purified mitochondrial NAD(H)-dependent GDH isoenzyme from soybean.     

Responses of the Circadian Locomotor Activity Rhythm of Mus Booduga to Shifts in LD Schedules

The responses of the field mouse Mus booduga to shifts in schedules of LD cycles were monitored and the results were interpreted with the help of a PRC constructed for the same species. The results reveal that, M. booduga reentrained faster with a lesser number of transients after delay shifts than advance shifts, thus exhibiting “asymmetry effect.” A positive correlation was observed between the number of transients and the number of hours of shift. In most of the shifts, the sign of the transients (negative for delaying transients and positive for advancing transients) coincided with the direction of the shift. Interestingly, 11 and 12 h of advance shifting resulted in delaying transients. An 11-h advance shift can also be interpreted as a 13-h delay. Reentrainment through delaying transients is faster as compared to reentrainment through advancing transients. Thus, this animal might have taken a “shorter route,” as proved by the fact that an 11-h advance shift has evoked delaying transients. But a 13-h advance shift evoked only advancing transients. This prompts us to speculate that there may be a “phase jump” in M. booduga. Further, irrespective of whether L or D has been doubled in a 12-h shift, both evoked only delaying transients.     

Molecular Cloning and Characterization of a Calmodulin Gene from Arabidopsis thaliana

The calcium binding protein, calmodulin Is involved in regulating various cellular and biochemical processes. A gene tor calmodulin (CaM) has been Isolated from a genomic library of Arabidopsis thaliana constructed in ; EMBL-4 using a heterologous cDNA probe from electric eel. One of the positive clones was characterized and the region containing the calmodulin gene sequences was Identified, excised using appropriate restriction enzymes and subcloned Into a plasmid vector. The genomic clone contains a complete copy of the calmodulin gene. A comparison of the nucleotide sequence of the part of the clone with those of the other plant and animal systems confirms that the clone In fact contains the calmodulin gene sequences. Southern hybridization ulling the calmodulin gene sequences as a probe reveals the presence of more than one copy of the calmodulin gene. The results of this investigation taken together with those Iff the other. indicate that the calmodulin gene belongs to a small mutigene family consisting of atieast four member. In the Arabidopsis genome.     

Mode of Dinitroaniline Herbicide Action: II. CHARACTERIZATION OF [C]ORYZALIN UPTAKE AND BINDING

The intracellular binding of dinitroaniline herbicides was studied in order to analyze the mechanism of their colchicine-like action. When corn root apices (5 millimeters) are incubated in [¹⁴C]oryzalin (a dinitroaniline herbicide), the ¹⁴C is taken up rapidly, reaching a plateau in about 4 hours, which corresponds to the minimum incubation time in oryzalin required to get maximum inhibition of elongation. At 4 hours, the [¹⁴C]oryzalin concentration inside the roots is 35 times higher than that in the incubation medium. Since this accumulation of [¹⁴C]oryzalin is not affected by 1 millimolar sodium azide and there is no metabolism of [¹⁴C]oryzalin under these conditions, the [¹⁴C]oryzalin must be accumulated (bound) in corn root apices by a process not requiring metabolic energy.     

Parasites,predators and other arthropods associated withMusca domestica and other flies breeding in bovine manure

The species of flies breeding in bovine manure, their parasites and predators as well as other associated arthropods in 3 localities in and near Bangalore are recoreded. The flies areMusca domestica L.,Musca pattoni Austen,Stomoxys calcitrans L.,Physiphora aenea F.,Physiphora demandata F.,Sargus metallinus F.,Sepsis thoracica R.-D.,Sepsis nitens Wiedemann,Sphaerocera scabricula Hal, andLeptocera (Coproica) hirtula Rondani. Four species of hymenopterous pupal parasites of these flies have been obtained:Spalangia cameroni Perkins from pupae ofM. domestica, M. pattoni, S. calcitrans, P. aenea andP. demandata; Spalangia endius Walker from pupae ofM. domestica, M. pattoni andS. calcitrans; Spalangia nigroaenea Curtis from pupae ofM. domestica andS. calcitrans; andDirhinus trichiophthalmus Masi from pupae ofSargus metallinus F. Percentage parasitism of fly puparia in field samples was noted. Four species ofHister andAleochara puberula Klug feeding on various immature stages ofM. domestica, 2 species of scarabaeid beetles, 2 species of ants and a pseudoscorpion were also found in the manure. The importance of biotic regulatory factors in the control of flies is discussed.

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Résumé Dans 3 localités de Bangalore ou proches de cette ville, on a inventorié les espèces de mouches vivant dans le fumier de vache, leurs parasites et prédateurs ainsi que d'autres arthropodes associés. Les mouches récoltées sont:Musca domestica L.,Musca pattoni Austen,Stomoxys calcitrans L.,Physiphora aenea F.,Physiphora demandata F.,Sargus metallinus F.,Sepsis thoracica R-D.,Sepsis nitens Wiedemann,Sphaerocera scabricula Hal. etLeptocera (Coproica) hirtula Rondani. Quatre espèces d'hyménoptères parasites des pupes ont été obtenues de ces mouches:Spalangia cameroni Perkins deM. domestica, M. pattoni, S. calcitrans, P. aenea etP. demandata: Spalangia endius Walker deM. domestica, M. pattoni etS. calcitrans, P. aenea etP. demandata; Spalangia endius Walker trans; etDirhinus trichiophthalmus Masi deSargus metallinus F. On a noté le taux de parasitisme des nymphes dans les conditions naturelles. Quatre espèces deHister etAleochara puberula Klug s'attaquant aux divers stades non imaginaux deM. domestica, 2 espèces de scarabeides, 2 espèces de fourmis et un pseudoscorpion ont été également trouvés dans le fumier. L'importance de ces facteurs biotiques dans la régulation des populations de mouches est discutée.

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Paper presented at the seminar on utilization of farm wastes for rural industrial growth, National Dairy Research Institute, Bangalore, 31 st December, 1975.

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This paper is published with the permission of the Director-General, Indian Council of Medical Research, New Delhi