Biochemical variation in the Sunni Muslims of Pulwama district, Jammu and Kashmir

Baseline data are presented on phenotype and gene frequency distributions of nine red-cell enzyme systems (ADA, AK, EsD, GLO I, PGM1, AP, GPI, PGM2, SOD) in the Sunni Muslims of Kashmir valley in the northernmost Indian border state of Jammu and Kashmir.     

A fertile revertant from petaloid cytoplasmic male-sterile carrot has a rearranged mitochondrial genome

 A spontaneously derived fertile plant was recovered from a petaloid cytoplasmic male-sterile (CMS) carrot inbred line. Genetic analysis indicated a single nuclear gene was responsible for the restoration to fertility. Within a family segregating for the nuclear restorer in combination with the sterility-inducing cytoplasm, fertile plants were recovered that could not restore fertility when crossed to sterile genotypes. Genetic analysis indicated cytoplasmic reversion for fertility, and Southern analysis, comparing mtDNA organization of the fertile revertant and its CMS progenitor, identified mitochondrial genome rearrangements. Hybridization of cosmids representing a 108-kb subgenomic circle of the sterile line to DNA of a fertile maintainer and fertile revertant lines indicated a similar mtDNA organization for these genotypes that was distinct from that of the sterile line. Six restriction fragments totalling 43.2 kb were common to the fertile maintainer and revertant and absent in the sterile; other restriction fragments totalling 38.2 kb were present only for the sterile line. Unique fragments of low stoichiometry, two for the fertile maintainer and three for the revertant, distinguished these lines. The reversion to fertility in the sterile line could have resulted from the amplification of a mitochondrial submolar genome highly homologous to that found in the fertile maintainer line. Received: 4 October 1997/Accepted: 12 December 1997     

Genetic epidemiology of non-insulin-dependent diabetes mellitus in north India: preliminary analyses of some genetic markers in Punjabis

Studies on monozygotic (MZ) twins and admixed populations show that the predisposition to non-insulin-dependent diabetes mellitus has a large genetic component. We have examined the distribution of some genetic polymorphisms (ABO, GLO, ESD, AK, ACPA, and GPI) in control and diabetic Punjabis from north India. The distribution of various genetic markers indicate that the differences between the control and diabetic samples are statistically not significant. Moreover, a contingency chi-square analysis over all loci suggests nonsignificant genetic differentiation (p = 0.50) between the Punjabi samples.     

Manufacturing of recombinant adeno‐associated viruses using mammalian expression platforms

Manufacturing practices for recombinant adeno‐associated viruses (AAV) have improved in the last decade through the development of new platforms in conjunction with better production and purification methods. In this review, we discuss the advantages and limitations of the most popular systems and methods employed with mammalian cell platforms. Methods and systems such as transient transfection, packaging and producer cells and adenovirus and herpes simplex virus are described. In terms of best production yields, they are comparable with about 10⁴–10⁵ vector genomes produced per cell but transient transfection of HEK293 cells is by far the most commonly used. For small‐scale productions, AAV can be directly purified from the producing cell lysate by ultracentrifugation on a CsCl or iodixanol‐step gradient whereas large‐scale purification requires a combination of multiple steps. Micro/macrofiltration (i.e. including tangential flow filtration and/or dead‐end filtration) and chromatography based‐methods are used for large‐scale purification. Purified AAV products must then be quantified and characterized to ensure quality. Recent purification methods and current analytical techniques are reviewed here. Finally, AAV technology is very promising, but manufacturing improvements are still required to meet the needs of affordable, safe and effective AAV vectors essential for licensing of gene therapy clinical protocols.     

Breath-by-breath measurement of the volume displaced by diaphragm motion.

To develop an accurate method to measure the volume displaced by diaphragm motion (DeltaVdi) breath by breath, we compared DeltaVdi measured by a previously evaluated biplanar radiographic method (Singh B, Eastwood PR, and Finucane KE. J Appl Physiol 91: 1913-1923, 2001) at several lung volumes during vital capacity inspirations in 10 healthy and nine hyperinflated subjects with 1) DeltaVdi measured from the same chest X-rays by two previously described uniplanar methods (Petroll WM, Knight H, and Rochester DF. J Appl Physiol 69: 2175-2182, 1990; Verschakelen JA, Deschepper K, and Demendts M. J Appl Physiol 72: 1536-1540, 1992) and a proposed method that considered actual cross-sectional shape of the rib cage and spinal volume (DeltaVdi(S)); and 2) DeltaVdi(S) measured by lateral fluoroscopy in the same 10 healthy subjects. Relative to biplanar DeltaVdi, DeltaVdi(S) values from lateral chest X-rays and fluoroscopy were not different, whereas DeltaVdi values of Petroll et al. and Verschakelen et al. were increased by (means +/- SD) 1.98 +/- 1.59 and 1.16 +/- 0.82 liters, respectively (both P < 0.001). During quiet breathing, DeltaVdi(S) by lateral fluoroscopy was 66 +/- 16% of tidal volume and similar to that between functional residual capacity and one-half inspiratory capacity by the biplanar radiographic method. We conclude that accurate breath-by-breath measurements of DeltaVdi can be made by using lateral fluoroscopy.     

Validation of a high-performance liquid chromatographic assay for the quantification of adenovirus type 5 particles

An anion-exchange–high-performance liquid chromatography (AE–HPLC) method for the quantification of adenovirus type 5 (Ad5) total particles was validated according to performance criteria of precision, specificity, linearity of calibration and range, limit of detection, limit of quantification, accuracy and recovery. The viral particles were detected by absorbance at 260 nm using photodiode array detector (PDA). Cesium chloride (CsCl) purified Ad5 and lysate samples were used for the validation of the method. Relative standard deviations (RSDs) for the inter-day, intra-day precision and reproducibility for both the lysate and the Ad5 standard were less than 10 and 2% for the peak area and retention time, respectively. The method was specific for Ad5 which was eluted at 8.0 min. The presence of DNA does not affect the recovery of Ad5 particles for accurate quantification. Based on the error in prediction to be less than 10%, the working range was established between 2×10¹⁰ and 7×10¹¹ VP/ml with correlation coefficient of 0.99975, standard deviation of 6.14×10⁹ VP/ml and a slope of 3.04×10⁵ VP/ml. The recovery of the method varied between 88 and 106% in all of the lysate samples investigated which is statistically similar to 100% recovery at 95% confidence interval.     

Uptake and translocation of zinc absorbed through roots and fruiting organs in peanuts

Peanut plants (Arachis hypogaea L.) are known to absorb Ca, P and S through the fruiting organs, but information on Zn uptake pattern is lacking. Therefore, a green-house experiment was conducted to study the uptake and translocation of Zn when applied in the rooting and fruiting zones of peanut plants. To locate the pathway and distribution of radioactive Zn, autoradiographs of plants were also taken.Zinc uptake data and autoradiographs indicated that a substantial amount of⁶⁵Zn was absorbed through the fruiting organs (auxillary system). Of the total⁶⁵Zn in the whole plant, 55.2 per cent was absorbed through roots and remaining 44.8 per cent through fruiting organs. Zinc was translocated to all the plant parts regardless of its absorption through roots or fruiting organs. The highest zinc concentration was recorded in the kernels, followed by leaves, stem and the shell.Contribution from the Department of Soils, Haryana Agric. Univ., Hissar-India.     

Limitations of conventional regression analysis a proposed modification

Summary The conventional genotype-environment interaction analysis cannot detect the theoretically ideal genotype which has been defined as the one with relatively low sensitivity in the poor environments and high sensitivity in the favourable environments. The computation of separate regression coefficients on the two regions of the response curve has been suggested to detect such genotypes. This procedure is simple and more convenient than the complicated curvilinear regression analysis.     

Repeatability of stability estimators for downy mildew incidence in pearl millet

Summary Repeatability of mean downy mildew (Sclerospora graminicola (Sacc.) Schroet.) incidence, regression coefficients and deviation mean squares were investigated for 25 pearl millet (Pennisetum typhoides (Burm.) Stapf. & Hubb.) genotypes in 20 environments by correlating arrays of these stability parameters over subsets of the 20 environments arranged according to the year-wise, random, stratified and extreme methods of environmental division. Correlation coefficients between arrays of mean downy mildew incidence from different pairs of subsets ranged from 0.57 to 0.98 and those of deviation mean squares from 0.58 to 0.96 indicating good repeatability of these parameters. Arrays of regression coefficients from different subsets, on the other hand, showed correlation coefficients that ranged from –0.58 to 0.96. Apparently, the regression index of stability was not repeatable for the genotypes and environments studied. Therefore, in order to identify a widely adapted genotype, testing is required to be carried out over a wider range of environments.     

Degradation of Alachlor [2-chloro-N-(methoxymethyl)-2′,6′-diethylacetanilide] by soil fungi

Summary A Penicillium sp. and a Trichoderma sp. were isolated from a soil previously treated with alachlor which is commonly used as herbicide. These fungi were found to degrade alachlor and only one degradation product was observed after 15 days of incubation. Whereas two products were noticed after 30 days with both the test organisms. re]19740913